Mechanisms of KCC2 upregulation during development

Gamma-aminobutyric acid (GABA) acting through ionotropic GABAA receptors plays a crucial role in the activity of the central nervous system (CNS). It triggers Ca2+ rise providing trophic support in developing neurons and conducts fast inhibitory function in mature neuronal networks. There is a developmental change in the GABAA reversal potential towards more negative levels during the first two postnatal weeks in rodent hippocampus. This change provides the basis for mature GABAergic activity and is attributable to the developmental expression of the neuron-specific potassium chloride cotransporter 2 (KCC2). In this work we have studied the mechanisms responsible for the control of KCC2 developmental expression. As a model system we used hippocampal dissociated cultures plated from embryonic day (E) 17 mice embryos before the onset of KCC2 expression. We showed that KCC2 was significantly up-regulated during the first two weeks of culture development. Interestingly, the level of KCC2 upregulation was not altered by chronic pharmacological blockage of action potentials as well as GABAergic and glutamatergic synaptic transmission. By in silico analysis of the proximal KCC2 promoter region we identified 10 candidate transcription factor binding sites that are highly conserved in mammalian KCC2 genes. One of these transcription factors, namely early growth response factor 4 (Egr4), had similar developmental profile as KCC2 and considerably increased the activity of mouse KCC2 gene in neuronal cells. Next we investigated the involvement of neurotrophic factors in regulation of Egr4 and KCC2 expression. We found that in immature hippocampal cultures Egr4 and KCC2 levels were strongly up-regulated by brain derived neurotrophic factor (BDNF)and neurturin. The effect of neurotrophic factors was dependent on the activation of a mitogen activated protein kinase (MAPK) signal transduction pathway. Intact Egr4-binding site in proximal KCC2 promoter was required for BDNF-induced KCC2 transcription. In vitro data were confirmed by several in vivo experiments where we detected an upregulation of KCC2 protein levels after intrahippocampal administration of BDNF or neurturin. Importantly, a MAPK-dependent rise in Egr4 and KCC2 expression levels was also observed after a period of kainic acid-induced seizure activity in neonatal rats suggesting that neuronal activity might be involved in Egr4-mediated regulation of KCC2 expression. Finally we demonstrated that the mammalian KCC2 gene (alias Slc12a5) generated two neuron-specific isoforms by using alternative promoters and first exons. A novel isoform of KCC2, termed KCC2a, differed from the previously known KCC2b isoform by 40 unique N-terminal amino acid residues. KCC2a expression was restricted to CNS,remained relatively constant during postnatal development, and contributed 20 50% of total KCC2 mRNA expression in the neonatal mouse brainstem and spinal cord. In summary, our data provide insight into the complex regulation of KCC2 expression during early postnatal development. Although basal KCC2 expression seems to be intrinsically regulated, it can be further augmented by neurotrophic factors or by enhanced activity triggering MAPK phosphorylation and Egr4 induction. Additional KCC2a isoform, regulated by another promoter, provides basal KCC2 level in neonatal brainstem and spinal cord required for survival of KCC2b knockout mice.

Characterization of genomic diversity in extraintestinal pathogenic Escherichia coli (ExPEC) and development of a diagnostic DNA microarray for the differentiation of ExPEC isolates causing urinary tract infections

Extraintestinal pathogenic Escherichia coli (ExPEC) represent a diverse group of strains of E. coli, which infect extraintestinal sites, such as the urinary tract, the bloodstream, the meninges, the peritoneal cavity, and the lungs. Urinary tract infections (UTIs) caused by uropathogenic E. coli (UPEC), the major subgroup of ExPEC, are among the most prevalent microbial diseases world wide and a substantial burden for public health care systems. UTIs are responsible for serious morbidity and mortality in the elderly, in young children, and in immune-compromised and hospitalized patients. ExPEC strains are different, both from genetic and clinical perspectives, from commensal E. coli strains belonging to the normal intestinal flora and from intestinal pathogenic E. coli strains causing diarrhea. ExPEC strains are characterized by a broad range of alternate virulence factors, such as adhesins, toxins, and iron accumulation systems. Unlike diarrheagenic E. coli, whose distinctive virulence determinants evoke characteristic diarrheagenic symptoms and signs, ExPEC strains are exceedingly heterogeneous and are known to possess no specific virulence factors or a set of factors, which are obligatory for the infection of a certain extraintestinal site (e. g. the urinary tract). The ExPEC genomes are highly diverse mosaic structures in permanent flux. These strains have obtained a significant amount of DNA (predictably up to 25% of the genomes) through acquisition of foreign DNA from diverse related or non-related donor species by lateral transfer of mobile genetic elements, including pathogenicity islands (PAIs), plasmids, phages, transposons, and insertion elements. The ability of ExPEC strains to cause disease is mainly derived from this horizontally acquired gene pool; the extragenous DNA facilitates rapid adaptation of the pathogen to changing conditions and hence the extent of the spectrum of sites that can be infected. However, neither the amount of unique DNA in different ExPEC strains (or UPEC strains) nor the mechanisms lying behind the observed genomic mobility are known. Due to this extreme heterogeneity of the UPEC and ExPEC populations in general, the routine surveillance of ExPEC is exceedingly difficult. In this project, we presented a novel virulence gene algorithm (VGA) for the estimation of the extraintestinal virulence potential (VP, pathogenicity risk) of clinically relevant ExPECs and fecal E. coli isolates. The VGA was based on a DNA microarray specific for the ExPEC phenotype (ExPEC pathoarray). This array contained 77 DNA probes homologous with known (e.g. adhesion factors, iron accumulation systems, and toxins) and putative (e.g. genes predictably involved in adhesion, iron uptake, or in metabolic functions) ExPEC virulence determinants. In total, 25 of DNA probes homologous with known virulence factors and 36 of DNA probes representing putative extraintestinal virulence determinants were found at significantly higher frequency in virulent ExPEC isolates than in commensal E. coli strains. We showed that the ExPEC pathoarray and the VGA could be readily used for the differentiation of highly virulent ExPECs both from less virulent ExPEC clones and from commensal E. coli strains as well. Implementing the VGA in a group of unknown ExPECs (n=53) and fecal E. coli isolates (n=37), 83% of strains were correctly identified as extraintestinal virulent or commensal E. coli. Conversely, 15% of clinical ExPECs and 19% of fecal E. coli strains failed to raster into their respective pathogenic and non-pathogenic groups. Clinical data and virulence gene profiles of these strains warranted the estimated VPs; UPEC strains with atypically low risk-ratios were largely isolated from patients with certain medical history, including diabetes mellitus or catheterization, or from elderly patients. In addition, fecal E. coli strains with VPs characteristic for ExPEC were shown to represent the diagnostically important fraction of resident strains of the gut flora with a high potential of causing extraintestinal infections. Interestingly, a large fraction of DNA probes associated with the ExPEC phenotype corresponded to novel DNA sequences without any known function in UTIs and thus represented new genetic markers for the extraintestinal virulence. These DNA probes included unknown DNA sequences originating from the genomic subtractions of four clinical ExPEC isolates as well as from five novel cosmid sequences identified in the UPEC strains HE300 and JS299. The characterized cosmid sequences (pJS332, pJS448, pJS666, pJS700, and pJS706) revealed complex modular DNA structures with known and unknown DNA fragments arranged in a puzzle-like manner and integrated into the common E. coli genomic backbone. Furthermore, cosmid pJS332 of the UPEC strain HE300, which carried a chromosomal virulence gene cluster (iroBCDEN) encoding the salmochelin siderophore system, was shown to be part of a transmissible plasmid of Salmonella enterica. Taken together, the results of this project pointed towards the assumptions that first, (i) homologous recombination, even within coding genes, contributes to the observed mosaicism of ExPEC genomes and secondly, (ii) besides en block transfer of large DNA regions (e.g. chromosomal PAIs) also rearrangements of small DNA modules provide a means of genomic plasticity. The data presented in this project supplemented previous whole genome sequencing projects of E. coli and indicated that each E. coli genome displays a unique assemblage of individual mosaic structures, which enable these strains to successfully colonize and infect different anatomical sites.

Expected Performance of the ATLAS Experiment - Detector, Trigger and Physics

A detailed study is presented of the expected performance of the ATLAS detector. The reconstruction of tracks, leptons, photons, missing energy and jets is investigated, together with the performance of b-tagging and the trigger. The physics potential for a variety of interesting physics processes, within the Standard Model and beyond, is examined. The study comprises a series of notes based on simulations of the detector and physics processes, with particular emphasis given to the data expected from the first years of operation of the LHC at CERN.

Zorros märke : Manligt identitetsbygge i Per Pettersons roman "Jeg forbanner tidens elv"

Avhandlingens syfte är att belysa hur porträtten av jagberättaren Arvid och hans mor växer fram i Per Pettersons roman Jeg forbanner tidens elv. Det paradigm jag utgår ifrån är det psykoanalytiska. Förutom texter av Sigmund Freud och Jacques Lacan stöder jag mig på texter av Peter Brooks och Terry Eagleton. För en tolkning av Arvids sätt att berätta sin historia använder jag mig av James Phelans tankar kring den opålitliga berättaren . Analysen bygger på en närläsning av romanen och några av de intertexter som förekommer i den. Av dessa är särskilt myten om kung Oidipus samt berättelsen om Zorro centrala för förståelsen av Arvids personlighet och hur den manliga identiteten byggs. Andra centrala intertexter som granskas närmare är Erich Maria Remarques roman Triumfbågen och Somerset Maughams roman Den vassa eggen. Porträttet av mor belyses indirekt via den funktion hon har i Arvids berättelse. Arvids porträtt analyseras ur två olika perspektiv. I uppsatsens första del, Romanbygget , undersöker jag hur romanen är uppbyggd och hur bilden av Arvid formas genom vad han berättar om sin mor, sitt liv, sin bakgrund och sina uppväxtår. Det perspektiv som Konung Oidipus i Freuds tolkning av det antika dramat ger, lyfter, som en nyckel in i romanen, fram dynamiken mellan Arvid, mor och den övriga familjen. I romanen dödas far i psykisk bemärkelse, han blir medvetet föraktad och förbisedd som manlig förebild och identifikationsobjekt. Arvids fixering vid mor gör att han ser sig själv med hennes ögon. Också bröderna får sin gestalt som rivaler i kampen om mor. I ljuset av den oidipala problematiken framhåller jag Arvids olösta relation till familjemedlemmarna som den avgörande orsaken till Arvids misslyckanden i livet och hans oförmåga att forma en fungerande och stabil vuxenidentitet. Men jag föreslår också en tidig, omedveten fadersidentifikation, symboliserad av Zorro och Zorros magiska märke, som i sublimeringen eventuellt finner sin lösning i en dröm om att bli författare. I uppsatsens andra del Berättarrösten undersöker jag Arvids sätt att berätta utgående ifrån Phelans tankar kring den opålitliga berättaren . Jag analyserar några centrala avsnitt i romanen med avseende på hur berättarröstens och den implicita författarens framställningar överensstämmer eller skiljer sig ifrån varandra. I min läsning är Arvid en komplext pålitlig och opålitlig berättare. Arvid framhåller i sin berättelse och i sina återblickar ett tillrättalagt och i någon mån förskönat porträtt av sig själv, en livslögn vars upplösning enligt min mening antyds i de avslutande kapitlen. För min förståelse av psykoanalysens teori och hur den kan tillämpas i litteraturforskningen är Ludwig Wittgensteins tankar om bildens användning centrala. I avsnittet om Zorro tar jag kortfattat upp frågan hur psykoanalytisk litteraturtolkning kan leda vilse i form av övertolkning, det vill säga att analysen övergår i fantasi. En annan möjlig felkälla som jag lyfter fram i analysen är att romanen tolkas av en svenskspråkig läsare som eventuellt läser in andra nyanser i den norska texten, än vad författaren avsett. Jag tar också upp frågan om Arvid i Pettersons tidigare produktion och huruvida det är frågan om en fortgående berättelse om Arvid Jansen under olika livsbetingelser. Mitt intryck är att det inte är fråga om ett enhetligt personporträtt utan olika frågeställningar som modelleras ur samma material.

Sapientia, 1975, Vol. XXX, nº 118 (número completo)

Contenido: Imago Dei / Octavio N. Derisi – El resultado de la actitud científica de Pascal / Jorge E. Saltor – Sobre la prueba ontólogica cartesiana / Agustín Seguí – Santo Tomás y Wittgenstein / Oscar S. Rotella – El primer argumento de la exposición metafísica del concepto del tiempo en la “Crítica de la razón pura” / Carlos A. Aldanondo – Acotaciones al problema de un aquende y un allende el conocimiento racional – Ivo Höllhuber – Notas y comentarios -- Bibliografía

Anuario de la Facultad de Ciencias Económicas del Rosario, Vol. 10, 2014

Contenido: Contabilidad y estados contables : implicancias del nuevo ordenamiento del Código Civil y Comercial de la Nación / Guillermo Boggino ; Clide Palacios -- De la inconmensurabilidad a la multimodalidad, un camino sin cisuras / Francisco Casiello ; Luis Herrera -- Ejercicios con la relación entre crecimiento económico y gasto público / Juan Sebastián Landoni -- La vigencia de las ideas de Ludwig Von Mises, para la interpretación de los problemas de Latinoamérica, en la actualidad / Guillermo Covernton -- Servicio de consulta / Patricia Constantini -- La inserción laboral de jóvenes profesionales con discapacidad en el marco de la responsabilidad social empresaria y los indicadores no financieros en la gestión sostenible / Analía Cinalli -- Una nueva menesterosidad para habitar la enseñanza de la filosofía / María del Huerto Revaz -- Los activos intangibles : una incertidumbre en la información contable / Vanesa Vazzano -- El modelo de gestión estratégica de recursos humanos como herramienta de desarrollo, crecimiento, eficiencia y modernización de la gestión pública / Hilda Delaux -- Estudio de la estabilidad de una variante del modelo de la telaraña / José Semitiel ; Angélica Arnulfo ; Cintia Cianciardo -- Microcréditos : tensión social ente destinatarios y tasa de interés / Germán Messina

The Real-Time Mach-Zehnder Interferometer Used In Space Experiment

The optical interference method is a promising technique for measuring temperature, density, and concentration in fluids. The non-intrusive and non-invasive nature of its optical techniques to the measured section are its most outstanding features. However, the adverse experiment environment, especially regarding shaking and vibrating, greatly restricts the application of the interferometer. In the present work, an optical diagnostic system consisting of a Mach-Zehnder interferometer (named after physicists Ludwig Mach) and an image processor has been developed that increases the measuring sensitivity compared to conventional experimental methods in fluid mechanics. An image processor has also been developed for obtaining quantitative results by using Fourier transformation. The present facility has been used in observing and measuring the mass transfer process of a water droplet in EAFP protein solution under microgravity condition provided by the satellite Shi Jian No. 8.

Prudentia Iuris, 2000, n° 52 (número completo)

Contenido: Editorial : veinte años de la revista fundada por el Dr. Santiago de Estrada – Derecho Eclesiástico: Condición jurídica del Estado de la Ciudad del Vaticano / Ariel David Busso – Derecho Comercial: La empresa y las responsabilidades emergentes de la competencia desleal / Isabel Miguez – Presentación en concurso preventivo de una (ex) entidad financiera / Javier Armando Lorente – La contenciosidad del procedimiento concursal / Claudio A. Galli – Panorama de la cooptación en el directorio privado en el Derecho Comparado / Mariano Gagliardo – Sobre los fundamentos: Teoría política de la costumbre y de la codificación : Montesquieu y la doble cultura jurídica en la antigua Francia / Paul-Ludwig Weinacht (traducción del Prof. Dr. Hugo Laitenberger) Universidad de Würzburg – Las certezas del sentido común y los caracteres esenciales de la pena / Mauro Ronco. Universidad de Modena, traducción de Alfredo Battaglia y comentario de Luis María Rizzi: “La pena como retribución y redención” – In memoriam – Documentos: Portada, índice y primer editorial del Nº 1 de “Prudentia Juris” en agosto de 1980 -- Recensiones

Legalidad y derecho natural institucional

Resumen: En su obra The morality of law, Lon Fuller pretende indicar ocho principios de la legalidad (principles of legality) que orientan la actividad de la legislación. La legalidad se presenta como un ejemplo de una concepción procedimental o institucional del derecho natural (procedure natural law o institutional natural law). El objeto del “iusnaturalismo institucional” de Fuller no es el contenido ético universal del derecho (derecho natural sustantivo), sino su modo formal-procedimental de existencia (derecho natural institucional). Este artículo tiene cuatro partes. Primero, el proyecto del iusnaturalismo institucional; segundo, los ocho principios de la legalidad de Fuller señalando su presencia también en las obras de Tomás de Aquino, Hobbes y Von Hayek; tercero, algunos conceptos de Wittgenstein y de John Searle que iluminan el aspecto metodológico del iusnaturalismo institucional y cuarto, se apuntará que el proyecto de un iusnaturalismo institucional ya está presente en la reflexión tomista sobre el ius gentium. En la conclusión, se recurrirá a las contribuciones de Jeremy Waldron para establecer el locus del iusnaturalismo institucional en el interior de la tradición iusnaturalista.

La burbuja inmobiliaria americana

Resumen: Axel Leijonhufvud, economista reconocido internacionalmente por sus trabajos sobre la literatura de John Maynard Keynes y el keynesianismo (Leijonhufvud, 1968), ofreció su lectura de la crisis subprime de 2008, reconociendo que la misma encaja más con la teoría austríaca del ciclo económico de Ludwig von Mises y Friedrich Hayek, que con el marco teórico keynesiano. En este artículo ofrecemos evidencia para tal afirmación.

El viajero Wilhelm von Humboldt y sus sucesores: caracterización cultural del País Vasco en las descripciones de viaje del siglo XIX

Berta Raposo Fernández e Ingrid García Wistädt (editoras)

Diário da Constituinte [gravação de vídeo] : [programa n. 676 1ª ed.]

Não houve quórum para votação dos Direitos Sociais, com apenas 234 parlamentares. Foram sugeridas propostas para ampliar a presença no plenário, tais como: chamar os suplentes e suspensos os ausentes sistemáticos e, ou não se pague ao constituinte que não aparecer. Foram sugeridas criações de Comissões que cuidem de grupos de emendas por assunto, para facilitar a votação, pois não haverá atrasos com as negociações. O PSDB reuniu líderes dos partidos para discutir os item mais polêmicos dos direitos sociais.

Diário da Constituinte [gravação de vídeo] : [programa n. 676A]

Debate entre constituintes e hematologistas quanto à definição de competências do Estado brasileiro no tocante à coleta, ao processamento e à transfusão de sangue e derivados. Inexistência de quorum regimental para realização de sessão. Divergência quanto ao impacto das eleições municipais sobre o andamento dos trabalhos constituintes.