Interaction of Mycoplasma genitalium with host cells: evidence for nuclear localization

Mycoplasma genitalium (Mg) is a mollicute that causes a range of human urogenital infections. A hallmark of these bacteria is their ability to establish chronic infections that can persist despite completion of appropriate antibiotic therapies and intact and functional immune systems. Intimate adherence and surface colonization of mycoplasmas to host cells are important pathogenic features. However, their facultative intracellular nature is poorly understood, partly due to difficulties in developing and standardizing cellular interaction model systems. Here, we characterize growth and invasion properties of two Mg strains (G37 and 1019V). Mg G37 is a high-passage laboratory strain, while Mg 1019V is a low-passage isolate recovered from the cervix. The two strains diverge partially in gene sequences for adherence-related proteins and exhibit subtle variations in their axenic growth. However, with both strains and consistent with our previous studies, a subset of adherent Mg organisms invade host cells and exhibit perinuclear targeting. Remarkably, intranuclear localization of Mg proteins is observed, which occurred as early as 30 min after infection. Mg strains deficient in adherence were markedly reduced in their ability to invade and associate with perinuclear and nuclear sites.

Slower rescue of ER homeostasis by the unfolded protein response pathway associated with common variable immunodeficiency

Common variable immunodeficiency (CVID) is a primary immunodeficiency characterized by hypogammaglobulinemia and recurrent infections. Herein we addressed the role of unfolded protein response (UPR) in the pathogenesis of the disease. Augmented unspliced X-box binding protein 1 (XBP-1) mRNA concurrent with co-localization of IgM and BiP/GRP78 were found in one CVID patient. At confocal microscopy analysis this patient`s cells were enlarged and failed to present the typical surface distribution of IgM, which accumulated within an abnormally expanded endoplasmic reticulum. Sequencing did not reveal any mutation on XBP-1, neither on IRE-1 alpha that could potentially prevent the splicing to occur. Analysis of spliced XBP-1, IRE-1 alpha and BiP messages after LPS or Brefeldin A treatment showed that, unlike healthy controls that respond to these endoplasmic reticulum (ER) stressors by presenting waves of transcription of these three genes, this patient`s cells presented lower rates of transcription, not reaching the same level of response of healthy subjects even after 48 h of ER stress. Treatment with DMSO rescued IgM and IgG secretion as well as the expression of spliced XBP-1. Our findings associate diminished splicing of XBP-1 mRNA with accumulation of IgM within the ER and lower rates of chaperone transcription, therefore providing a mechanism to explain the observed hypogammaglobulinemia. (C) 2008 Elsevier Ltd. All rights reserved.

Strong surjectivity of maps from 2-complexes into the 2-sphere

Given a model 2-complex K(P) of a group presentation P, we associate to it an integer matrix Delta(P) and we prove that a cellular map f : K(P) -> S(2) is root free (is not strongly surjective) if and only if the diophantine linear system Delta(P) Y = (deg) over right arrow (f) has an integer solution, here (deg) over right arrow (f) is the so-called vector-degree of f

Global analytic regularity for structures of co-rank one

We consider real analytic involutive structures V, of co-rank one, defined on a real analytic paracompact orientable manifold M. To each such structure we associate certain connected subsets of M which we call the level sets of V. We prove that analytic regularity propagates along them. With a further assumption on the level sets of V we characterize the global analytic hypoellipticity of a differential operator naturally associated to V. As an application we study a case of tube structures.

Semantic mapping using mobile robots

Robotic mapping is the process of automatically constructing an environment representation using mobile robots. We address the problem of semantic mapping, which consists of using mobile robots to create maps that represent not only metric occupancy but also other properties of the environment. Specifically, we develop techniques to build maps that represent activity and navigability of the environment. Our approach to semantic mapping is to combine machine learning techniques with standard mapping algorithms. Supervised learning methods are used to automatically associate properties of space to the desired classification patterns. We present two methods, the first based on hidden Markov models and the second on support vector machines. Both approaches have been tested and experimentally validated in two problem domains: terrain mapping and activity-based mapping.

Theoretical Analysis of Aggregation in Block-Copolymer Films: The Optical Signature

We focus this work on the theoretical investigation of the block-copolymer poly [oxyoctyleneoxy-(2,6-dimethoxy-1,4phenylene-1,2-ethinylene-phenanthrene-2,4diyl) named as LaPPS19, recently proposed for optoelectronic applications. We used for that a variety of methods, from molecular mechanics to quantum semiempirical techniques (AMI, ZINDO/S-CIS). Our results show that as expected isolated LaPPS19 chains present relevant electron localization over the phenanthrene group. We found, however, that LaPPS19 could assemble in a pi-stacked form, leading to impressive interchain interaction; the stacking induces electronic delocalization between neighbor chains and introduces new states below the phenanthrene-related absorption; these results allowed us to associate the red-shift of the absorption edge, seen in the experimental results, to spontaneous pi-stack aggregation of the chains. (C) 2009 Wiley Periodicals, Inc. Int J Quantum Chem 110: 885-892, 2010

Ligand induced interaction of thyroid hormone receptor beta with its coregulators

Thyroid hormones exert most of their physiological effects through two thyroid hormone receptor (TR) subtypes, TR alpha and TR beta, which associate with many transcriptional coregulators to mediate activation or repression of target genes. The search for selective TR beta ligands has been stimulated by the finding that several pharmacological actions mediated by TR beta might be beneficial in medical conditions such as obesity, hypercholesterolemia and diabetes. Here, we present a new methodology which employs surface plasmon resonance to investigate the interactions between TR beta ligand binding domain (LBD) complexes and peptides derived from the nuclear receptor interaction motifs of two of its coregulators, SRC2 and DAX1. The effect of several TR beta ligands, including the TR beta selective agonist GC-I and the TR beta selective antagonist NH-3, were investigated. We also determined the kinetic rate constants for the interaction of TR beta-T3 with both coregulators, and accessed the thermodynamic parameters for the interaction with DAX1. Our findings Suggest that flexibility plays an important role in the interaction between the receptor and its coregulators. and point out important aspects of experimental design that should be addressed when using TR beta LBD and its agonists. Furthermore, the methodology described here may be useful for the identification of new TR beta ligands. (C) 2008 Elsevier Ltd. All rights reserved.

Cathepsin X cleaves the beta 2 cytoplasmic tail of LFA-1 inducing the intermediate affinity form of LFA-1 and alpha-actinin-1 binding

The motility of T cells depends on the dynamic spatial regulation of integrin-mediated adhesion and de-adhesion. Cathepsin X, a cysteine protease, has been shown to regulate T-cell migration by interaction with lymphocyte function associated antigen-1 (LFA-1). LFA-1 adhesion to the ICAM-1 is controlled by the association of actin-binding proteins with the cytoplasmic tail of the beta(2) chain of LFA-1. Cleavage by cathepsin X of the amino acid residues S(769), E(768) and A(767) from the C-terminal of the beta(2) cytoplasmic tail of LFA-1 is shown to promote binding of the actin-binding protein alpha-actinin-1. Furthermore, cathepsin X overexpression reduced LFA-1 clustering and induced an intermediate affinity LFA-1 conformation that is known to associate with a-actinin-1. increased levels of intermediate affinity LFA-1 resulted in augmented cell spreading due to reduced attachment of T cells to the ICAM-1-coated surface. Gradual cleavage of LFA-1 by cathepsin X enables the transition between intermediate and high affinity LFA-1, an event that is crucial for effective T-cell migration.

Associated Family of G-Structure Preserving Minimal Immersions in Semi-Riemannian Manifolds

We prove the existence of an associated family of G-structure preserving minimal immersions into semi-Riemannian manifolds endowed with a compatible infinitesimally homogeneous G-structure. We will study in more details minimal embeddings into product of space forms.

Metabotropic glutamate receptors transduce signals for neurite outgrowth after binding of the prion protein to laminin gamma 1 chain

The prion protein (PrP(C)) is highly expressed in the nervous system, and its abnormal conformer is associated with prion diseases. PrP(C) is anchored to cell membranes by glycosylphosphatidylinositol, and transmembrane proteins are likely required for PrP(C)-mediated intracellular signaling. Binding of laminin (Ln) to PrP(C) modulates neuronal plasticity and memory. We addressed signaling pathways triggered by PrP(C)-Ln interaction in order to identify transmembrane proteins involved in the transduction of PrP(C)-Ln signals. The Ln gamma 1-chain peptide, which contains the Ln binding site for PrP(C), induced neuritogenesis through activation of phospholipase C (PLC), Ca(2+) mobilization from intracellular stores, and protein kinase C and extracellular signal-regulated kinase (ERK1/2) activation in primary cultures of neurons from wild-type, but not PrP(C)-null mice. Phage display, coimmunoprecipitation, and colocalization experiments showed that group I metabotropic glutamate receptors (mGluR1/5) associate with PrP(C). Expression of either mGluR1 or mGluR5 in HEK293 cells reconstituted the signaling pathways mediated by PrP(C)-Ln gamma 1 peptide interaction. Specific inhibitors of these receptors impaired PrP(C)-Ln gamma 1 peptide-induced signaling and neuritogenesis. These data show that group I mGluRs are involved in the transduction of cellular signals triggered by PrP(C)-Ln, and they support the notion that PrP(C) participates in the assembly of multiprotein complexes with physiological functions on neurons.-Beraldo, F. H., Arantes, C. P., Santos, T. G., Machado, C. F., Roffe, M., Hajj, G. N., Lee, K. S., Magalhaes, A. C., Caetano, F. A., Mancini, G. L., Lopes, M. H., Americo, T. A., Magdesian, M. H., Ferguson, S. S. G., Linden, R., Prado, M. A. M., Martins, V. R. Metabotropic glutamate receptors trans-duce signals for neurite outgrowth after binding of the prion protein to laminin gamma 1 chain. FASEB J. 25, 265-279 (2011). www.fasebj.org

Sdo1p, the yeast orthologue of Shwachman-Bodian-Diamond syndrome protein, binds RNA and interacts with nuclear rRNA-processing factors

The Shwachman-Bodian-Diamond syndrome protein (SBDS) is a member of a highly conserved protein family of not well understood function, with putative orthologues found in different organisms ranging from Archaea, yeast and plants to vertebrate animals. The yeast orthologue of SBDS, Sdo1p, has been previously identified in association with the 60S ribosomal subunit and is proposed to participate in ribosomal recycling. Here we show that Sdo1p interacts with nucleolar rRNA processing factors and ribosomal proteins, indicating that it might bind the pre-60S complex and remain associated with it during processing and transport to the cytoplasm. Corroborating the protein interaction data, Sdo1p localizes to the nucleus and cytoplasm and co-immunoprecipitates precursors of 60S and 40S subunits, as well as the mature rRNAs. Sdo1p binds RNA directly, suggesting that it may associate with the ribosomal subunits also through RNA interaction. Copyright (C) 2009 John Wiley & Sons, Ltd.

"La leyenda tiene una base real" : Análisis crítico del discurso de un artículo de El País

This paper carries out a critical discourse analysis (CDA) of the article "¿Chinofobia?” published by the Spanish newspaper El País. It applies the theory of racist discourse in the media as formulated by Teun A. van Dijk. Its hypothesis is that the article, which supposedly analyses discrimination against the Chinese minority in Spain, covertly blames the Chinese minority itself for the discrimination. Applying CDA methodology as exemplified in previous studies by van Dijk, the paper analyses the article on global and local levels, and delineates its mental model. The global level analysis describes the article in terms of macropropositions and proposes a macrostructure: the self-alienation of the Chinese minority. The paper then analyses how the macrostructure is reinforced on the local level through micropropositions by examining 1) how vocabulary serves the negative presentation and othering of the Chinese minority, 2) how strategies of mitigation minimise discrimination by employing imprecise and vague language, 3) how quotes are used to give coherence and force to the macrostructure, 4) how implications associate the Chinese minority with criminality, and 5) how stereotyped beliefs about the Chinese minority are presented as common sense (presuppositions) and fallaciously argued. Finally, the paper delineates the mental model: the presuppositions about integration, and the implicit warning that minorities should integrate or they will be discriminated against.

High-growth firms : Not so vital after all?

High-growth firms have received considerable interest recently since they create most of the new jobs in the economy. The purpose of our paper is to investigate the characteristics of high-growth firms prior to their growth period, and whether these characteristics differ across industries. Using data on a large sample of limited liability firms in Sweden for the period 2007-2010, we find that high-growth firms do not have the characteristics that we typically associate with successful firms. On the contrary, our results indicate that high-growth firms have low profits and a weak financial position. This might explain why studies have found that high-growth firms are seldom capable of sustaining their high growth rates in subsequent periods, and thus question policies that are targeted towards these companies.

Can humming caregivers´ facilitate feeding during mealtime situations with persons with dementia? A qualitative study

Background. ‘Music Therapeutic Caregiving’, defined as when caregivers’ sing for or together with persons with dementia care situations, has been shown to facilitate the caring situation, and enhance positive and decrease negative expressions in persons with dementia. No studies about singing during mealtimes have been conducted, and this project was designed to elucidate this. However, since previous studies have shown that there is a risk that persons with dementia will start to sing along with the caregiver, the caregiver in this study hummed such that the person with dementia did not sing instead of eat. Aim. To describe professional caregivers’ experiences of humming during meal time while feeding persons with dementia. Method. An intervention with caregivers humming was implemented during lunch time. Focus group interviews were conducted to fetch the caregivers experiences. Qualitative content analysis was used to analyse the interviews. Results. The caregivers described that humming disseminated relaxation and joy, and awakened memories about the songs hummed which encouraged the persons with the dementia to join in the songs. They also described that humming seemed to make the persons with dementia associate with mealtime and could make them eat more. However it also revealed suspiciousness and agitation from the persons with dementia. Conclusion. Humming during mealtime revealed mainly positive as well as some negative influences. More and larger studies are needed to be able to draw general conclusions.

Towards a Formalisation of Electronic Contracting Environments

The IST-CONTRACT project is in the process of creating an electronic contracting language. One of the goals of this language is that it has formal underpinnings, and formalizations at a number of levels have been created. One of the lowest levels, upon which the other levels are built is the normative level. At this level, we identify how contract clauses (modeled as norms) may evolve over time. In this paper, we describe this formalization, and show how we may associate various states with a norm throughout its lifecycle. We also show how more complex evaluations may be carried out over a norm, and conclude with an example showing the application of the framework over a contract and its associated norms.