Virulence properties of Aggregatibacter actinomycetemcomitans biofilm and characterisation of its putative cytokine exploitation

Biofilms are surface-attached multispecies microbial communities that are embedded by their self-produced extracellular polymeric substances. This lifestyle enhances the survival of the bacteria and plays a major role in many chronic bacterial infections. For instance, periodontitis is initiated by multispecies biofilms. The phases of active periodontal tissue destruction and notably increased levels of proinflammatory mediators, such as the key inflammatory mediator interleukin (IL)-1beta, are typical of the disease. The opportunistic periodontal pathogen Aggregatibacter actinomycetemcomitans is usually abundant at sites of aggressive periodontitis. Despite potent host immune system responses to subgingival invaders, A. actinomycetemcomitans is able to resist clearance attempts. Moreover, some strains of A. actinomycetemcomitans can generate genetic diversity through natural transformation, which may improve the species’ adjustment tothe subgingival environment in the long term. Some biofilm forming species are known to bind and sense human cytokines. As a response to cytokines, bacteria may increase biofilm formation and alter their expression of virulence genes. Specific outer membrane receptors for interferon-γ or IL-1β have been characterised in two Gram-negative pathogens. Because little is known about periodontal pathogens’ ability to sense cytokines, we used A. actinomycetemcomitans as a model organism to investigate how the species responds to IL-1beta. The main aims of this thesis were to explore cytokine binding on single-species A. actinomycetemcomitans biofilms and to determine the effects of cytokines on the biofilm formation and metabolic activity of the species. Additionally, the cytokine’s putative internalisation and interaction with A. actinomycetemcomitans proteins were studied. The possible impact of biofilm IL-1beta sequestering on the proliferation and apoptosis of gingival keratinocyte cells was evaluated in an organotypic mucosa co-culture model. Finally, the role of the extramembranous domain of the outer membrane protein HofQ (emHofQ) in DNA binding linked to DNA uptake in A. actinomycetemcomitans was examined. Our main finding revealed that viable A. actinomycetemcomitans biofilms can bind and take up the IL-1β produced by gingival cells. At the sites of pathogen-host interaction, the proliferation and apoptosis of gingival keratinocytes decreased slightly. Notably, the exposure of biofilms to IL-1beta caused their metabolic activity to drop, which may be linked to the observed interaction of IL-1beta with the conserved intracellular proteins DNA binding protein HU and the trimeric form of ATP synthase subunit beta. A Pasteurellaceaespecific lipoprotein, which had no previously determined function, was characterized as an IL-1beta interacting membrane protein that was expressed in the biofilm cultures of all tested A. actinomycetemcomitans strains. The use of a subcellular localisation tool combined with experimental analyses suggested that the identified lipoprotein, bacterial interleukin receptor I (BilRI), may be associated with the outer membrane with a portion of the protein oriented towards the external milieu. The results of the emHofQ study indicated that emHofQ has both the structural and functional capability to bind DNA. This result implies that emHofQ plays a role in DNA assimilation. The results from the current study also demonstrate that the Gram-negative oral species appears to sense the central proinflammatory mediator IL-1beta.

Resistência de populações de capim-colchão (Digitaria ciliaris) aos herbicidas inibidores da Acetil Co-A Carboxilase

O objetivo do presente trabalho foi avaliar as populações com suspeita de ocorrência de biótipos resistentes de Digitaria ciliaris aos herbicidas inibidores da AcetilCoA Carboxilase (ACCase), por meio de curvas de dose-resposta, bem como estabelecer o grau de resistência cruzada aos herbicidas cicloexanodionas (CHD) e ariloxifenoxipropionatos (APP) desses biótipos. O experimento foi conduzido em casa de vegetação, utilizando-se quatro populações com suspeita de resistência (R1, R2, R3 e R4) e uma população suscetível (S). O delineamento experimental adotado foi o de blocos ao acaso, com quatro repetições, sendo os tratamentos resultado da interação fatorial entre cinco populações, três herbicidas (fluazifop-p-butil, sethoxydim e tepraloxydim) e oito doses de herbicidas (0C, 0,06C, 0,125C, 0,5C, 1C, 2C, 4C e 10C), em que C é a dose comercial recomendada para cada produto. Foram realizadas avaliações de porcentagem de controle aos 28 dias após a aplicação (DAA). A partir dos resultados obtidos, conclui-se que as populações R1, R2, R3 e R4 apresentaram-se como biótipos resistentes aos herbicidas inibidores da ACCase, com diferentes níveis de resistência cruzada aos herbicidas com esse mecanismo de ação.

Vesicular Trafficking in Osteoclasts

Osteoclasts are multinucleated bone-degrading cells that undergo large changes in their polarisation and vesicular trafficking during the bone resorption cycle. Rab proteins are small GTPases that offer both temporal and spatial regulation to the transport between membranous organelles. Previously the presence and function of only few of the currently known 60 Rab proteins in osteoclasts have been reported. In this study, the expression of 26 Rab genes in bone-resorbing osteoclasts was demonstrated with gene-specific primer pairs. The further analysis of three Rab genes during human osteoclast differentiation revealed that Rab13 gene is highly induced during osteoclastogenesis. The presence of Rab13 protein in the secretory vesicles directed towards the ruffled border and in the endocytotic or transcytotic pathways in resorbing osteoclasts was excluded. The localisation of Rab13 suggests that that it is associated with a previously unknown vesicle population travelling between the trans-Golgi network and the basolateral membrane in bone resorbing osteoclasts. Rab proteins convey their functions by binding to specific effector proteins. We found a novel Rab13 interaction with endospanins-1 and -2 that are yet poorly characterised small transmembrane proteins. The Rab13 subfamily member Rab8 also bound to endospanins, while Rab10 and unrelated Rabs did not. Rab13 and endospanin-2 co-localised in perinuclear vesicles in transfected cells, demonstrating the interaction also in vivo. The inhibition of Rab13 did not interfere with the localisation of endospanin-2 nor did it affect the cell surface expression of growth hormone receptor, as has been previously described for endospanins. The physiological role of this novel protein-protein interaction thus remains to be clarified. The analysis of the transcytotic route in bone resorbing osteoclasts revealed that multiple vesicle populations arise from the ruffled border and transport the bone degradation products for exocytosis. These vesicles are directed to the functional secretory domain that is encircled by an actin-based molecular barrier. Furthermore, the transcytotic vesicles contain abundant Helix pomatia lectin binding sites and represent lipid raft concentrates. Finally, autophagosomal compartments may also be involved in the transcytosis.

Value co-creation in gaming the industry: A qualitative research on virtual discussion forums

This study discusses the procedures of value co-creation that persist in gaming industry. The purpose of this study was to identify the procedures that persist in current video gaming industry which answers the main research problem how value is co-created in video gaming industry followed by three sub questions: (i) What is value co-creation in gaming industry? (ii) Who participates in value co-creation in gaming industry? (iii) What are the procedures that are involved in value co-creation in gaming industry? The theoretical background of the study consists of literature relating to the theory of marketing i.e., notion of value, conventional understanding of value creation, value chain, co-creation approach, co-production approach. The research adopted qualitative research approach. As a platform of relationship researcher used web 2.0 tool interface. Data were collected from the social networks and netnography method was applied for analyzing them. Findings show that customer and company both co-create optimum level of value while they interact with each other and within the customers as well. However mostly the C2C interaction, discussions and dialogues threads that emerged around the main discussion facilitated to co-create value. In this manner, companies require exploiting and further motivating, developing and supporting the interactions between customers participating in value creation. Hierarchy of value co-creation processes is the result derived from the identified challenges of value co-creation approach and discussion forums data analysis. Overall three general sets and seven topics were found that explored the phenomenon of customer to customer (C2C) and business to customer (B2C) interaction/debating for value co-creation through user generated contents. These topics describe how gamer contributes and interacts in co-creating value along with companies. A methodical quest in current research literature acknowledged numerous evolving flows of value in this study. These are general management perspective, new product development and innovation, virtual customer environment, service science and service dominant logic. Overall the topics deliver various realistic and conceptual implications for using and handling gamers in social networks for augmenting customers’ value co-creation process.

Pour les recherches sur les côtes occidentales de l'Afrique de Ptolemée et sur les isles connues par les ancien dans l'Océan Atlantique ; Côtes occidentales de l'Afrique selon Ptolemée

Julkaisussa: Recherches sur la géographie systématique et positive des anciens pour servir de base à l'histoire de la géographie ancienne. Vol I

Chiamasi Rossia tutto quel tratto, il qual da fonti del fiume Vistula confina cól Monte Carpato, col fiume Tira, e col Boristine fino a Kiovia, et indi, abbracciádo la puicia Seuvera ...

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan

Questa e la perfetta et vera descrittione et il vero disegno et sito del Mare Adriatico, di Larcipelago, del Mare di Soria, et del Mare Egeo, con li nomi volgari che hoggidi Seusano da Italiani, de l'Isole, Scogli, Capi, Golfi, Porti, et luochi che in ditti mari se ritruovano à o de l'Isole quanto delle infrascritte Prouicie di tema ferma, cioè di Bytinia, Misia, Licia, e Cilicia, del Peloponneso detto Morea, de l'Albania, et della Dalmatia, con la maggior parte dell'Italia, descritte secondo che in le carte da navigar se co[n]tegono

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan

Poly-DL-lactide-co-glycolide microspheres as a controlled release antigen delivery system

Successful vaccine application means maximum protection with minimal number of administrations. A rational development of vaccines involves studies of the nature of the antigen as well as of the adjuvant to be used to improve the immune responses. This has provided the impetus for studies to design the degradable devices and for different approaches to antigen delivery by different routes of administration. The development of controlled release systems based on polymeric devices that permit a sustained or pulsed release of encapsulated antigens has attracted much interest. Polymeric delivery systems consist of polymers that release their content continuously in a controlled manner over a period of time. The development of a biocompatible delivery system for parenteral administration offers several advantages in terms of immunoadjuvanticity over other compounds. It was found that, in contrast to other carriers, microspheres are more stable, thus permitting administration by the oral or parenteral route. In the present study, we describe the main characteristics and potentialities of this new immunoadjuvant for oral and parenteral administration.

Polen in die dermaligen Besitzungen eingetheilt : nach der vortreflichen Karte = Carte de la Pologne divisée entre les puissances co-partageantes

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan

Co-creating value with customers as a new approach of entering new market

Global trends associated with development of information technology, globalization, industrial and economic changes are influencing on company and customer domains and thus transforming company-customer relationship. The company centric paradigm with a strong product focus shifts to a customer oriented one with a strong emphasis on customer collaboration. As a result, the customer role changes from a passive observer to an active player. Moreover, global trends contribute to transformation of competitive environment making it tougher and simplifying an access to resources previously considered as unique. All that factors push the companies towards cooperation with customers in order to identify unarticulated needs and finding the best possible solution to existing customer problems. The Master’s Thesis is done for Outotec (Lappeenranta) which considers extension of dewatering business in Russian coal market. Research aims to identify key features of coal preparation and dewatering of fine coal and tailings in Russian preparation plants; analyze the state of Russian coal market and evaluate market potential for Outotec dewatering solutions. The study has a qualitative nature and implements an action research methodology that involves both creation of knowledge and introduction of changes into the system. The base for taking actions is formed by theoretical framework that targets on describing company - customer interaction and has selected co-creation as the most appropriate method of customer involvement. The integration of co-creation approach into an action research cycle allows not only fulfilling the research objectives but also facilitates organizational learning and intraorganizational collaboration, assists in establishing customer contacts and making the first steps into the market, bringing new joint projects to the company and opening real business opportunities.

Modulation of intercellular communication by differential regulation and heteromeric mixing of co-expressed connexins

Intercellular communication may be regulated by the differential expression of subunit gap junction proteins (connexins) which form channels with differing gating and permeability properties. Endothelial cells express three different connexins (connexin37, connexin40, and connexin43) in vivo. To study the differential regulation of expression and synthesis of connexin37 and connexin43, we used cultured bovine aortic endothelial cells which contain these two connexins in vitro. RNA blots demonstrated discordant expression of these two connexins during growth to confluency. RNA blots and immunoblots showed that levels of these connexins were modulated by treatment of cultures with transforming growth factor-ß1. To examine the potential ability of these connexins to form heteromeric channels (containing different connexins within the same hemi-channel), we stably transfected connexin43-containing normal rat kidney (NRK) cells with connexin37 or connexin40. In the transfected cells, both connexin proteins were abundantly produced and localized in identical distributions as detected by immunofluorescence. Double whole-cell patch-clamp studies showed that co-expressing cells exhibited unitary channel conductances and gating characteristics that could not be explained by hemi-channels formed of either connexin alone. These observations suggest that these connexins can readily mix with connexin43 to form heteromeric channels and that the intercellular communication between cells is determined not only by the properties of individual connexins, but also by the interactions of those connexins to form heteromeric channels with novel properties. Furthermore, modulation of levels of the co-expressed connexins during cell proliferation or by cytokines may alter the relative abundance of different heteromeric combinations.

A geological and topographical sketch map of the New York and Westmoreland Gas Coal Company's lands, at Manor Sta., Westm'd Co., Pa.

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan

Mer des Indes : côte occidentale de l'Hindoustan de Bombay à Calicut : Archipel des Laquedives

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan