Genetic and Molecular Basis of Encapsulation and Capsule Diversity in Kingella kingae

Kingella kingae is a bacterial pathogen that is increasingly recognized as an etiology of septic arthritis, osteomyelitis, bacteremia, and endocarditis in young children. The pathogenesis of K. kingae disease starts with bacterial adherence to the respiratory epithelium of the posterior pharynx. Previous work has identified type IV pili and a trimeric autotransporter protein called Knh (Kingella NhhA homolog) as critical factors for adherence to human epithelial cells. Additional studies established that the presence of a polysaccharide capsule interferes with Knh-mediated adherence. Given the inhibitory role of capsule during adherence we sought to uncover the genes involved in capsule expression to understand how capsule is elaborated on the cell surface. Additionally, this work aimed to further characterize capsule diversity among K. kingae clinical isolates and to investigate the relationship between capsule type and site of isolation.

We first set out to identify the carbohydrates present in the K. kingae capsule present in the prototype strain 269-492. Glycosyl composition and NMR analysis of surface extractable polysaccharides demonstrated two distinct polysaccharides, one consisting of GalNAc and Kdo with the structure →3)-β-GalpNAc-(1→5)-β-Kdop-(2→ and the other containing galactose alone with the structure →5)-β-Galf-(1→.

To discern the two polysaccharides we disrupted the ctrA gene required for surface localization of the K. kingae polysaccharide capsule and observed a loss of GalNAc and Kdo but no effect on the presence of Gal in bacterial surface extracts. In contrast, deletion of the pamABCDE locus involved in production of a reported galactan exopolysaccharide eliminated Gal but had no effect on the presence of GalNAc and Kdo in surface extracts. These results established that K. kingae strain KK01 produces a polysaccharide capsule with the structure →3)-β-GalpNAc-(1→5)-β-Kdop-(2→ and a separate exopolysaccharide with the structure →5)-β-Galf-(1→.

Having established that K. kingae produces a capsule comprised of GalNAc and Kdo, we next set out to identify the genetic determinants of capsule through a transposon mutagenesis screen. In addition to the previously identified ctrABCD operon, lipA, lipB, and a putative glycosyltransferase termed csaA (capsule synthesis region A gene A) were found to be essential for the production of surface-localized capsule. The ctr operon, lipA, lipB, and csaA were found to be present at unlinked locations throughout the genome, which is atypical for gram-negative organisms that elaborate a capsule dependent on an ABC-type transporter for surface localization. Through examining capsule localization in the ctrA, lipA, lipB, and csaA mutant strains, we determined that the ctrABCD, lipA/lipB, and csaA gene products respectively function in capsule export, assembly, and synthesis, respectively. The GalNAc transferase and Kdo transferase domains found in CsaA further support its role in catalyzing the synthesis of the GalNAc-Kdo capsule in the K. kingae prototype strain.

To investigate the capsule diversity that exists in K. kingae we screened a panel of strains isolated from patients with invasive disease or healthy carriers for the csaA capsule synthesis locus. We discovered that Kingella kingae expresses one of 4 capsule synthesis loci (csa, csb, csc, or csd) associated with a capsule consisting of Kdo and GalNAc (type a), Kdo and GlcNAc (type b), Kdo and ribose (type c), and GlcNAc and galactose (type d), respectively. Cloning of the csa, csb, csc, or csd locus into the empty flanking gene region in a non-encapsulated mutant (creation of an isogenic capsule swap) was sufficient to produce either the type a, type b, or type c capsule, respectively, further supporting the role of these loci in expression of a specific polysaccharide linkage. Capsule type a and capsule type b accounted for 96% of invasive strains. Conversely, capsule type c and capsule type d were found disproportionately among carrier isolates, suggesting that capsule type is important in promoting invasion and dissemination.

In conclusion, we discovered that Kingella kingae expresses a polysaccharide capsule and an exopolysaccharide on its surface that require distinct genetic loci for surface localization. Further investigation into genetic determinants of encapsulation revealed the loci ctrABCD, lipA/lipB, and a putative glycosyltransferase are required for capsule expression, with the gene products having roles in capsule export, assembly, and synthesis, respectively. The putative glycosyltransferase CsaA was determined to be a bifunctional enzyme with both GalNAc-transferase and Kdo-transferase activity. Furthermore, we discovered a total of 4 capsule types expressed in clinical isolates of K. kingae, each with a distinct capsule synthesis locus. The variation in the proportion of capsule types found between invasive strains and carriage strains suggest that capsule type is important in promoting invasion and dissemination. Taken together, this work expands our knowledge of the capsule types expressed among K. kingae carrier and invasive isolates and provides insights into the common genetic determinants of capsule expression. These contributions may lead to selecting clinically relevant capsule types to develop into a capsule based vaccine to prevent K. kingae colonization.

Standardization in the Pre-Columbian Pottery from El Caño, Panama: Specialization, Productivity and Consumption

The pottery found in the burials of El Cano is uniform in style to these made in the coclesanos valleys between 700 and 1000 AD. The coefficient of variability of the different pottery forms, evidence diverse standardizations values for polychrome and non-polychrome ceramics. Moreover, data of funerary contexts from the Cano recently excavated, suggest that elite has controlled ceramic production. This control over the production of certain goods reveals that these were important in the support or proper operational of the chiefdoms in Panama and mark the phase of splendour of this culture.

Petrofac International Limited: el mercadeo interno, una estrategia efectiva en el manejo de personal

Como futuros administrados es importante recordar que nuestra función no será únicamente crear una empresa y generar constantemente ingresos. Esto es una de tantas tareas que tiene una persona que se encamina por crear o ser parte de una empresa, sea grande o pequeña siempre existirán variables que con los años se han convertido más y más indispensables en los procesos de este recorrido. El tema principal de nuestro estudio de caso es entender el concepto del endomarketing o como muchos lo conocen marketing interno; en pocas palabras es algo que por años se ha visto su efectividad en la productividad y satisfacción de los empleados. Google es una de las multinacionales más grandes del mundo, que el ultimo reconocimiento fue por parte de uno de los ranking más influyentes “The World’s Billionaries” ocupando como el CEO de la compañía Larry Page el puesto #12 , que por años ha sabido invertir en su recurso más importante que son sus empleados. En una entrevista con la revista Fortune, Larry exclama que “es importante que la empresa sea una familia, que las personas sientan que son parte de la empresa, y que la empresa es como una familia para ellos. Cuando se trata a las personas de esa manera, se obtiene una mejor productividad”. Con este pequeño ejemplo e idea de cómo una de las marcas más influyentes en el mundo ha fortalecido su mejor recurso por años el cual le ha remunerado con el éxito que tiene hoy en día. Entonces es como el mercadeo interno o endomarketing siendo uno de los conceptos tratados en el mercadeo en general hoy en día, logrando meterse como un modelo de negocio en la mayoría de empresas que existen hoy. Este es uno de los motivos que nos motivó para desarrollar este estudio de caso que verán a continuación, el cual se basa en la investigación y análisis de variables que comprenden a la empresa Petrofac, y si bien ha sabido implementar el concepto en sus procesos diarios. El estudio de caso describe como a través de los años ha sabido implementar con éxito la herramienta de evaluación y análisis del mercado interno a los empleados que constituyen hoy en día Petrofac International Limited. Abarcaremos primero con los objetivos del estudio, seguido del inicio y crecimiento de la empresa en los últimos años, en que países tiene presencia y cuáles son sus proyectos más importantes hoy en día. El estudio de caso será implementado por un tipo de investigación descriptiva, que con la recopilación de suficiente información tanto cuantitativa como cualitativa nos ira fortaleciendo la hipótesis principal, usaremos datos estadísticos que serán recopilados de encuestas, entrevistas y focus group. Al final con el análisis y a las conclusiones que lleguemos se busca demostrar el impacto positivo que ha tenido los empleados de diferentes países y de diferentes culturas que poco a poco han influenciado a la empresa para alcanzar el éxito que ha logrado hasta ahora, y de la misma forma como la empresa ha influenciado en cada una de las vidas de estas personas sin importar el lugar en el que estén laborando, siempre la empresa busca que se sientan parte de ella. Como resultados generales, se obtienen argumentos que sustentan que la implementación del mercadeo interno le permite a la empresa generar ventajas competitivas y dinámicas, que son desarrolladas internamente para un futuro reflejarlas ante los clientes externos. El estudio permitió concluir que el mercadeo interno debe ser una estrategia de comunicación dentro de una empresa convirtiéndose de primera necesidad para la compañía, pues los beneficios que genera son cada vez más necesarios para cada área que abarca el negocio.

Warianty realizacji austriackości w wybranych autorów (Joseph Roth, Stefan Zweig, Franz Theodor Csokor i Ődön von Horváth) w odniesieniu do kategorii granicznych starego i nowego świata

Wydział Neofilologii:Instytut Filologii Germańskiej

Certificate that Joseph Kingsmill, sheriff, has sold 3 ½ acres in Lot no. 34 in the 3rd Concession of Wainfleet to W. H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 3 ½ acres in Lot no. 34 in the 3rd Concession of Wainfleet to W. H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 3 acres in Lot no. 7 [?] in Grimsby to W. H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 3 acres in Lot no. 7 [?] in Grimsby to W. H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 6 acres in Lot no. 9 in the 4th Concession of Caistor to W.H. Dickson [originally said S.D. Woodruff, but this was crossed out]

Certificate that Joseph Kingsmill, sheriff, has sold 6 acres in Lot no. 9 in the 4th Concession of Caistor to W.H. Dickson [originally said S.D. Woodruff, but this was crossed out], May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no. 18 in the 4th Concession of Humberland to W.H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no. 18 in the 4th Concession of Humberland to W.H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no.1 in the 6th Concession of Grimsby to W. H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no.1 in the 6th Concession of Grimsby to W. H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no.33 in the 4th Concession of Wainfleet to W.H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Lot no.33 in the 4th Concession of Wainfleet to W.H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 7 acres in Lot no. 24 in the 2nd Concession of Caistor to W. H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 7 acres in Lot no. 24 in the 2nd Concession of Caistor to W. H. Dickson, May 23, 1857.

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Broken Front Lot no.3 in the 3rd Concession of Gainboro to W.H. Dickson

Certificate that Joseph Kingsmill, sheriff, has sold 4 acres in Broken Front Lot no.3 in the 3rd Concession of Gainsboro to W.H. Dickson, May 23, 1857.

Questions and answers of W. Joseph Smith, clerk of Grimsby

Questions and answers of W. Joseph Smith, clerk of Grimsby (2 pages, handwritten). These pages are stained and slightly torn. This affects a small portion of the text, 1830.

Examination of W. Joseph Smith, clerk of Grimsby

Examination of W. Joseph Smith, clerk of Grimsby (2 pages, handwritten). The pages are stained and torn. This affects the text slightly, 1830.

Broadside of a requisition to W. Woodruff from William W. Ball, John McBride, Robert N. Ball, W. Servos, Joseph Wynn and 150 other people for Mr. Woodruff to run as a Representative in Parliament

Broadside, 45 cm. x 30 cm. of a requisition to W. Woodruff from William W. Ball, John McBride, Robert N. Ball, W. Servos, Joseph Wynn and 150 other people for Mr. Woodruff to run as a Representative in Parliament. This is followed by a positive, and humble response from Mr. Woodruff, June 3, 1856.