Interrogation of human monoclonal antibodies induced by 4C-MenB to identify protective antigens contained in the OMV component

Neisseria meningitidis is a gram negative human obligated pathogen, mostly found as a commensal in the oropharyngeal mucosa of healthy individuals. It can invade this epithelium determining rare but devastating and fast progressing outcomes, such as meningococcal meningitidis and septicemia, leading to death (about 135000 per year worldwide). Conjugated vaccines for serogroups A, C, W135, X and Y were developed, while for N. meningitidis serogroup B (MenB) the vaccines were based on Outern Membrane Vesicles (OMV). One of them is the 4C-MenB (Bexsero). The antigens included in this vaccine’s formulation are, in addition to the OMV from New Zeland epidemic strain 98/254, three recombinant proteins: NadA, NHBA and fHbp. While the role of these recombinant components was deeply characterized, the vesicular contribution in 4C-MenB elicited protection is mediated mainly by porin A and other unidentified antigens. To unravel the relative contribution of these different antigens in eliciting protective antibody responses, we isolated human monoclonal antibodies (mAbs) from single-cell sorted plasmablasts of 3 adult vaccinees peripheral blood. mAbs have been screened for binding to 4C-MenB components by Luminex bead-based assay. OMV-specific mAbs were purified and tested for functionality by serum bactericidal assay (SBA) on 18 different MenB strains and characterized in a protein microarray containing a panel of prioritized meningococcal proteins. The bactericidal mAbs identified to recognize the outer membrane proteins PorA and PorB, stating the importance of PorB in cross-strain protection. In addition, RmpM, BamE, Hyp1065 and ComL were found as immunogenic components of the 4C-MenB vaccine.

Outer membrane vesicles from cold-adapted antarctic bacteria

Many Gram-negative, cold-adapted bacteria from the Antarctic environment produce large amounts of extracellular matter with potential biotechnological applications. Transmission electron microscopy (TEM) analysis after high-pressure freezing and freeze substitution (HPF-FS) showed that this extracellular matter is structurally complex, appearing around cells as a netlike mesh, and composed of an exopolymeric substance (EPS) containing large numbers of outer membrane vesicles (OMVs). Isolation, purification and protein profiling via 1D SDS-PAGE confirmed the outer membrane origin of these Antarctic bacteria OMVs. In an initial attempt to elucidate the role of OMVs in cold-adapted strains of Gram-negative bacteria, a proteomic analysis demonstrated that they were highly enriched in outer membrane proteins and periplasmic proteins associated with nutrient processing and transport, suggesting that the OMVs may be involved in nutrient sensing and bacterial survival. OMVs from Gram-negative bacteria are known to play a role in lateral DNA transfer, but the presence of DNA in these vesicles has remained difficult to explain. A structural study of Shewanella vesiculosa M7T using TEM and Cryo-TEM revealed that this Antarctic Gram-negative bacterium naturally releases conventional one-bilayer OMVs, together with a more complex type of OMV, previously undescribed, which on formation drags along inner membrane and cytoplasmic content and can therefore also entrap DNA.

Outer membrane vesicles from cold-adapted antarctic bacteria

Many Gram-negative, cold-adapted bacteria from the Antarctic environment produce large amounts of extracellular matter with potential biotechnological applications. Transmission electron microscopy (TEM) analysis after high-pressure freezing and freeze substitution (HPF-FS) showed that this extracellular matter is structurally complex, appearing around cells as a netlike mesh, and composed of an exopolymeric substance (EPS) containing large numbers of outer membrane vesicles (OMVs). Isolation, purification and protein profiling via 1D SDS-PAGE confirmed the outer membrane origin of these Antarctic bacteria OMVs. In an initial attempt to elucidate the role of OMVs in cold-adapted strains of Gram-negative bacteria, a proteomic analysis demonstrated that they were highly enriched in outer membrane proteins and periplasmic proteins associated with nutrient processing and transport, suggesting that the OMVs may be involved in nutrient sensing and bacterial survival. OMVs from Gram-negative bacteria are known to play a role in lateral DNA transfer, but the presence of DNA in these vesicles has remained difficult to explain. A structural study of Shewanella vesiculosa M7T using TEM and Cryo-TEM revealed that this Antarctic Gram-negative bacterium naturally releases conventional one-bilayer OMVs, together with a more complex type of OMV, previously undescribed, which on formation drags along inner membrane and cytoplasmic content and can therefore also entrap DNA.

Prevalencia de enfermedad de Fabry en pacientes en lista de trasplante y posttrasplante renal en fundación cardioinfantil Bogotá

Introducción: La Enfermedad de Fabry (EF), es una enfermedad multisistémica de almacenamiento lisosomal ligada al cromosoma X que afecta principalmente a hombres, pero también puede causar significativa morbilidad en las mujeres heterocigotas (1–5). La deficiencia de la enzyma α-galactosidaseA (α-Gal A,) provoca acumulación de glicosfingolipidos que afectan diferentes tipos celulares entre ellos el endotelio vascular en vasos de pequeño calibre, células epiteliales y Músculo liso en el sistema cardiovascular (cardiomiocitos), sistema nervioso y células epiteliales tubulares del riñón (6,7). Complicaciones como la falla renal es la causa de muerte más frecuente en la EF (7,8). La incidencia se ha calculado en 1 de cada 117.000 nacidos vivos. (9). Objetivos: Determinar la prevalencia de la Enfermedad de Fabry en pacientes con Insuficiencia renal terminal que se encuentren en lista de trasplante y Post-trasplante Renal en Fundación Cardioinfantil Bogotá. Materiales y Métodos: Se realizó un estudio observacional en donde se evaluó la prevalencia de la EF en todos los sujetos mayores de 18 años que se encuentren en lista de trasplante y post-trasplante renal. Resultados: La prevalencia de Enfermedad de Fabry en 98 pacientes con enfermedad renal crónica fue de 7.1% para la muestra general y 12.9% para la muestra con etiología idiopática Conclusiones: La Enfermedad de Fabry es una importante casusa de Enfermedad Renal Crónica Terminal principalmente en el grupo de etiología idiopática. Palabras Clave: Enfermedad de Fabry (FA)

Klassifikation av vårdåtgärder, KVÅ

Bakgrund: Behovet av att göra verksamhetsuppföljningar som bland annat grundar sig på individrelaterade åtgärder är stort inom hälso- och sjukvård och omsorg. Inom omvårdnadsområdet finns ett antal klassifikationer främst framtagna i USA. Inom medicinska området finns specialitetsspecifika klassifikationer som till exempel Klassifikation av kirurgiska åtgärder. Någon patientrelaterad, vårdgivaroberoende klassifikation har emellertid inte funnits i Sverige. Socialstyrelsen påbörjade därför ett arbete att utveckla en svensk klassifikation, oberoende av vårdgivarkategori och medicinska specialiteter. Ett första förslag till klassifikation av medicinska åtgärder, KMÅ, presenterades år 2000. Förslaget ansågs inte representera en tillräcklig bredd av åtgärder varför Svensk sjuksköterskeförening, Förbundet Sveriges arbetsterapeuter och Legitimerade sjukgymnasters förbund år 2004 fick ett uppdrag att komplettera och utveckla denna klassifikation att också gälla andra vårdåtgärder än rent medicinska.Syfte: Att utveckla en tvärprofessionell klassifikation som beskriver patientrelaterade åtgärder inom omvårdnad, arbetsterapi och sjukgymnastik. Arbetet skulle inkluderas i en redan fastställd klassifikation med medicinsk inriktning.Metod: En arbetsgrupp bestående av sjuksköterskor, arbetsterapeuter och sjukgymnaster har utvecklat ett förslag till åtgärdsklassifikation – en testmodul - som utgår från komponenterna i WHO:s Klassifikation av funktionstillstånd, funktionshinder och hälsa, ICF. Valet av struktur grundar sig på att åtgärder inom arbetsterapi, omvårdnad och sjukgymnastik sällan har som fokus kroppsstruktur utan snarare individens funktioner, aktiviteter och delaktighet samt omgivningsfaktorer. Ett protokoll för prövning av testmodulen har utformats. Resultat: Socialstyrelsen antog arbetsgruppens förslag i november 2005. I december 2005 publicerade Socialstyrelsen förslaget i form av en testmodul som skall prövas bland annat i klinisk verksamhet under år 2006. Förslaget omfattar ca 250 centrala åtgärder på aggregerad nivå inom de tre verksamhetsområdena. Utifrån resultaten från prövningarna skall förslaget justeras och ytterligare utvecklas. Detta arbete beräknas vara klart under våren 2007 för att integreras till en gemensam tvärprofessionell åtgärdsklassifikation som kan tas i bruk årsskiftet 2007/2008.

Ultra-structural mapping of sugarcane bagasse after oxalic acid fiber expansion (OAFEX) and ethanol production by Candida shehatae and Saccharomyces cerevisiae

Background Diminishing supplies of fossil fuels and oil spills are rousing to explore the alternative sources of energy that can be produced from non-food/feed-based substrates. Due to its abundance, sugarcane bagasse (SB) could be a model substrate for the second-generation biofuel cellulosic ethanol. However, the efficient bioconversion of SB remains a challenge for the commercial production of cellulosic ethanol. We hypothesized that oxalic-acid-mediated thermochemical pretreatment (OAFEX) would overcome the native recalcitrance of SB by enhancing the cellulase amenability toward the embedded cellulosic microfibrils. Results OAFEX treatment revealed the solubilization of hemicellulose releasing sugars (12.56 g/l xylose and 1.85 g/l glucose), leaving cellulignin in an accessible form for enzymatic hydrolysis. The highest hydrolytic efficiency (66.51%) of cellulignin was achieved by enzymatic hydrolysis (Celluclast 1.5 L and Novozym 188). The ultrastructure characterization of SB using scanning electron microscopy (SEM), atomic force microscopy (AFM), Raman spectroscopy, Fourier transform–near infrared spectroscopy (FT-NIR), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) revealed structural differences before and after OAFEX treatment with enzymatic hydrolysis. Furthermore, fermentation mediated by C. shehatae UFMG HM52.2 and S. cerevisiae 174 showed fuel ethanol production from detoxified acid (3.2 g/l, yield 0.353 g/g; 0.52 g/l, yield, 0.246 g/g) and enzymatic hydrolysates (4.83 g/l, yield, 0.28 g/g; 6.6 g/l, yield 0.46 g/g). Conclusions OAFEX treatment revealed marked hemicellulose degradation, improving the cellulases’ ability to access the cellulignin and release fermentable sugars from the pretreated substrate. The ultrastructure of SB after OAFEX and enzymatic hydrolysis of cellulignin established thorough insights at the molecular level.

Use of E. coli OMVs as delivery system to elicit neutralizing antibodies against Chlamydia infectivity: the HtrA protein example

The obligate intracellular pathogen Chlamydia trachomatis is a gram negative bacterium which infects epithelial cells of the reproductive tract. C. trachomatis is the leading cause of bacterial sexually transmitted disease worldwide and a vaccine against this pathogen is highly needed. Many evidences suggest that both antigen specific-Th1 cells and antibodies may be important to provide protection against Chlamydia infection. In a previous study we have identified eight new Chlamydia antigens inducing CD4-Th1 and/or antibody responses that, when combined properly, can protect mice from Chlamydia infection. However, all selected recombinant antigens, upon immunization in mice, elicited antibodies not able to neutralize Chlamydia infectivity in vitro. With the aim to improve the quality of the immune response by inducing effective neutralizing antibodies, we used a novel delivery system based on the unique capacity of E. coli Outer Membrane Vesicles (OMV) to present membrane proteins in their natural composition and conformation. We have expressed Chlamydia antigens, previously identified as vaccine candidates, in the OMV system. Among all OMV preparations, the one expressing HtrA Chlamydia antigen (OMV-HtrA), showed to be the best in terms of yield and quantity of expressed protein, was used to produce mice immune sera to be tested in neutralization assay in vitro. We observed that OMV-HtrA elicited specific antibodies able to neutralize efficiently Chlamydia infection in vitro, indicating that the presentation of the antigens in their natural conformation is crucial to induce an effective immune response. This is one of the first examples in which antibodies directed against a new Chlamydia antigen, other than MOMP (the only so far known antigen inducing neutralizing antibodies), are able to block the Chlamydia infectivity in vitro. Finally, by performing an epitope mapping study, we investigated the specificity of the antibody response induced by the recombinant HtrA and by OMV-HtrA. In particular, we identified some linear epitopes exclusively recognized by antibodies raised with the OMV-HtrA system, detecting in this manner the antigen regions likely responsible of the neutralizing effect.

The NeOn Ontology Models

Interoperability on multiple levels, concerning both the ontologies themselves and their engineering activities, is a key requirement for ontology networks to be efficient, with minimal redundancy and high reuse. This requirement has a strict binding for software tools that can support some interoperability levels, yet they can be hindered by a lack of shared models and vocabularies describing the resources to be handled, as well as the ways of handling them. Here, three examples of metalevel vocabularies are proposed, each covering at least one peculiar interoperability aspect: OMV for modeling the artifacts themselves, LIR for managing a multilingual layer on top of them, and C-ODO Light for modeling collaboration-supportive life cycle management tasks and processes. All of these models lend themselves to handling by dedicated software tools and are all being employed within NeOn products.

Integración en la plataforma 4mobile de mecanismos de atención en redes sociales

El presente texto se ha desarrollado como Proyecto Fin de Grado en la Escuela Técnica Superior de Ingeniería y Sistemas de Telecomunicación de la Universidad Politécnica de Madrid en colaboración con la Consultora Tecnológica everis. El mismo tiene como objetivo realizar un estudio de los requisitos necesarios para poder desplegar un servicio que permita ofrecer a las Operadoras Móviles Virtuales (OMV) soporte técnico a sus clientes a través de las redes sociales. Este módulo establecerá una nueva vía de comunicación entre el consumidor y el cliente de la OMV con la propia OMV, el cual se caracterizará por una alta accesibilidad y una gran rapidez en el servicio permitiendo de este modo, una mayor satisfacción del cliente con su operador móvil y por tanto de una nueva vía para conseguir la fidelización del mismo y la captación de nuevos clientes. Para ello, este proyecto se lleva a cabo sobre el sistema de información 4mobile el cual es comercializado por la mencionada Consultora Tecnológica. Este sistema consiste en una plataforma web la cual permite cubrir todos los procesos de negocio comunes que un OMV necesita gestionar. Es por ello, que el mencionado estudio se centra en la evaluación de los aspectos necesarios para la integración de un módulo de estas características dentro de la plataforma de 4mobile. Este módulo, estará basado en una herramienta software que permitirá gestionar el ciclo de vida completo del comentario realizado por el cliente a través de una red social, desde que es publicado por el cliente, hasta que se haya respondido al mismo y su solución sea considerada como satisfactoria por el cliente. Por ello, y de cara a definir correctamente esta herramienta, será necesario un detallado análisis el cual recoja diversos aspectos y que a lo largo del texto, será denominado como Plan de Marketing de Medios Sociales (PMMS). Estos aspectos versarán tanto de las necesidades tecnológicas para su mencionada integración, como de la serie de características funcionales que una solución basada en servicio técnico a través de las redes sociales deberá poseer con el objetivo de ofrecer un servicio técnico de calidad. Finalmente, estas funcionalidades y necesidades tecnológicas se expondrán en forma de propuesta a everis para su integración en la plataforma 4mobile junto con un análisis de diseño a alto nivel software de la solución a desarrollar. ABSTRACT. This text has been developed as Final Degree Project in the Escuela Técnica Superior de Ingeniería y Sistemas de Telecomunicación de la Universidad Politécnica de Madrid in collaboration with the Technology Consultant everis, aims to conduct a study of how to provide technical support through social networks and the evaluation of the integration of a social support module within the system platform 4mobile, which cover all business processes that need to manage a Mobile Virtual Network Operator (MVNO), which is marketed by above-mentioned consultant . This module will establish a new communication channel between the consumer and the client with the OMV and itself, which is characterized by high accessibility and great fast service. Thus will allow a higher customer satisfaction with him service and thus a new way to get the same loyalty and attract new customers To this end, this project is performed on the information system 4mobile which is marketed by the consulting part thereof everis. This system is a platform to cover all business processes that need to manage an MVNO. Therefore, the mentioned study focuses on the evaluation of the elements necessary for integrating module these features within the platform 4mobile. This module will be based on a software tool for managing the entire lifecycle of the comment made by the customer via social networking, from the moment it is published, until it has been respond and their solution is considered satisfactory for the customer. So, in order to properly define this tool, a detailed analysis which will be necessary to collect various aspects and throughout the text, it will be referred to as Plan of Social Media Marketing (PMMS). These aspects will be address both: the technological needs for the mentioned integration, and the number of functional characteristics-based service through social networking solution, as this must in order to provide a quality service Finally, these goals and requirements will be discussed as everis offer for integration into the platform 4mobile analysis along with a high level of software design to develop the solution.

Nord Stream expansion: what does it mean for Europe? EPC Commentary, 2 July 2015

Two announcements in sequence have shaken the energy debate on 18 and 19 June. Gazprom announced the signature of a Memorandum of Understanding with Shell, Exon and OMV for the construction of strings 3 and 4 of Nord Stream, aiming at doubling the current 55 bcm capacity of the corridor running in the Baltic sea bed and connecting the Russian terminal of Vyborg to Germany. On the day after, a €2 bn deal between Russia and Greece was signed for extending the Turkish Stream project into Greek territory. ...

Nord Stream II – Another test for EU unity. EPC Commentary, 9 October 2015

In September 2015, Gazprom and major European energy companies (Germany's E.ON and BASF/Wintershall, Austria's OMV, France’s ENGIE and Royal Dutch Shell) signed an agreement to double the Nord Stream gas pipeline’s capacity by 2019. The plan is to transport up to 55 billion additional cubic meters of gas a year to Germany through the Baltic Sea, bypassing Ukraine and other Central-Eastern European countries.

Nord Stream II-testing EU and credibility. EPC Commentary, 16 December 2015

In September 2015, Gazprom and major European energy companies (Germany's E.ON and BASF/Wintershall, Austria's OMV, France’s ENGIE and Royal Dutch Shell) signed an agreement to double the Nord Stream gas pipeline’s capacity by 2019. The plan is to transport up to 55 billion additional cubic metres of gas a year to Germany through the Baltic Sea, bypassing Ukraine and other Central-Eastern European countries.

Energiapolitika – a 2008. év fejleményei

Az írás a hazai vezetékes energiapiacokat érintő legfontosabb tényleges 2008. évi fejleményeket vizsgálja. Kiemelt figyelmet szentel két témának: az év eleji árampiaci modellváltás értékelésének és az alternatív gázvezeték tervekről folytatott politikai vita elemzésének. A műhelytanulmány terjedelmi és hangsúlybéli okokból nem foglalkozik az Európai Uniós energiapolitikai fejlemények összefoglalásával és elemzésével, valamint a MOL-OMV párharc 2008. évi fejleményeivel.

Global distribution of modern shallow marine shorelines. Implications for exploration and reservoir analogue studies

Acknowledgments Support for this work came from the SAFARI consortium which was funded by Bayern Gas, ConocoPhillips, Dana Petroleum, Dong Energy, Eni Norge, GDF Suez, Idemitsu, Lundin, Noreco, OMV, Repsol, Rocksource, RWE, Statoil, Suncor, Total, PDO, VNG and the Norwegian Petroleum Directorate (NPD). This manuscript has benefited from discussion with Bruce Ainsworth, Rachel Nanson and Christian Haug Eide. Boyan Vakarelov and Richard Davis Jr. are thanked for their constructive reviews and valuable comments that helped to improve the manuscript.

Rotation of Vibrio fischeri Flagella Produces Outer Membrane Vesicles That Induce Host Development

Using the squid-vibrio association, we aimed to characterize the mechanism through which Vibrio fischeri cells signal morphogenesis of the symbiotic light-emitting organ. The symbiont releases two cell envelope molecules, peptidoglycan (PG) and lipopolysaccharide (LPS) that, within 12 h of light organ colonization, act in synergy to trigger normal tissue development. Recent work has shown that outer membrane vesicles (OMVs) produced by V. fischeri are sufficient to induce PG-dependent morphogenesis; however, the mechanism(s) of OMV release by these bacteria has not been described. Like several genera of both beneficial and pathogenic bacteria, V. fischeri cells elaborate polar flagella that are enclosed by an extension of the outer membrane, whose function remains unclear. Here, we present evidence that along with the well-recognized phenomenon of blebbing from the cell's surface, rotation of this sheathed flagellum also results in the release of OMVs. In addition, we demonstrate that most of the development-inducing LPS is associated with these OMVs and that the presence of the outer membrane protein OmpU but not the LPS O antigen on these OMVs is important in triggering normal host development. These results also present insights into a possible new mechanism of LPS release by pathogens with sheathed flagella. IMPORTANCE Determining the function(s) of sheathed flagella in bacteria has been challenging, because no known mutation results only in the loss of this outer membrane-derived casing. Nevertheless, the presence of a sheathed flagellum in such host-associated genera as Vibrio, Helicobacter, and Brucella has led to several proposed functions, including physical protection of the flagella and masking of their immunogenic flagellins. Using the squid-vibrio light organ symbiosis, we demonstrate another role, that of V. fischeri cells require rotating flagella to induce apoptotic cell death within surface epithelium, which is a normal step in the organ's development. Further, we present evidence that this rotation releases apoptosis-triggering lipopolysaccharide in the form of outer membrane vesicles. Such release may also occur by pathogens but with different outcomes for the host.