Enterohemorrhagic Escherichia coli colonization of human colonic epithelium in vitro and ex vivo

Enterohemorrhagic E. coli (EHEC) are important foodborne pathogens causing gastroenteritis and more severe complications such as hemorrhagic colitis and hemolytic uremic syndrome. Pathology is most pronounced in the colon, but to date there is no direct clinical evidence showing EHEC binding to colonic epithelium in patients. In this study, we investigated EHEC adherence to the human colon by using in vitro organ culture (IVOC) of colonic biopsies and polarized T84 colon carcinoma cells. We showed for the first time that EHEC colonized human colonic biopsies by forming typical attaching/effacing (A/E) lesions which were dependent on EHEC type III secretion (T3S) and binding of the outer membrane protein intimin to the Translocated intimin receptor (Tir). A/E lesion formation was dependent on oxygen levels and suppressed under oxygen-rich culture conditions routinely used for IVOC. In contrast, EHEC adherence to polarized T84 cells occurred independently of T3S and intimin and did not involve Tir translocation into the host cell membrane. Neither colonization of biopsies nor T84 cells was significantly affected by expression of Shiga toxins. Our study suggests that EHEC colonize and form stable A/E lesions on the human colon which is likely to contribute to intestinal pathology during infection. Furthermore, care needs to be taken when using cell culture models as they might not reflect the in vivo situation

Shiga toxin production and translocation during microaerobic human colonic infection with Shiga toxin-producing E. coli O157:H7 and O104:H4

Haemolytic uraemic syndrome caused by Shiga toxin-producing E. coli (STEC) is dependent on release of Shiga toxins (Stxs) during intestinal infection and subsequent absorption into the bloodstream. An understanding of Stx-related events in the human gut is limited due to lack of suitable experimental models. In this study, we have used a vertical diffusion chamber system with polarized human colon carcinoma cells to simulate the microaerobic (MA) environment in the human intestine and investigate its influence on Stx release and translocation during STEC O157:H7 and O104:H4 infection. Stx2 was the major toxin type released during infection. Whereas microaerobiosis significantly reduced bacterial growth as well as Stx production and release into the medium, Stx translocation across the epithelial monolayer was enhanced under MA versus aerobic conditions. Increased Stx transport was dependent on STEC infection and occurred via a transcellular pathway other than macropinocytosis. While MA conditions had a similar general effect on Stx release and absorption during infection with STEC O157:H7 and O104:H4, both serotypes showed considerable differences in colonization, Stx production, and Stx translocation which suggest alternative virulence strategies. Taken together, our study suggests that the MA environment in the human colon may modulate Stx-related events and enhance Stx absorption during STEC infection

Transposição de marcadores microssatélites derivados de mamona em tungue.

Entre as culturas oleaginosas, o tungue é uma alternativa de grande potencial econômico para o sul do Brasil por apresentar elevado rendimento de óleo. Embora genótipos introduzidos no Estado tenham demonstrado adaptação, é fundamental desenvolver um programa de melhoramento genético para a cultura, a fim de oferecer cultivares mais produtivas. Uma forma eficiente de auxiliar os programas de melhoramento é a análise da variabilidade genética por meio de marcadores moleculares. Muitos estudos têm mostrado que grande parte dos marcadores SSR encontrados numa espécie podem ser transferidos para espécies correlatas. Tanto a mamona (Ricinus communis L.) como o tungue (Aleurites fordii), pertencem à família Euphorbiaceae, o que pode facilitar a transposição de primers microssatélites de mamona para tungue. O presente trabalho teve como objetivo testar a transposição de marcadores microssatélites do genoma de mamona para tungue. Foram utilizados 74 pares de primers sintetizados a partir do genoma da mamona. Os resultados demonstram ser possível utilizar marcadores microssatélites em genótipos de tungue desenvolvidos a partir do genoma de mamona

Investigation of fiber/matrix adhesion: test speed and specimen shape effects in the cylinder test

The cylinder test, developed from the microdroplet test, was adapted to assess the interfacial adhesion strength between fiber and matrix. The sensitivity of cylinder test to pull-out speed and specimen geometry was measured. It was established that the effect of test speed can be described as a superposition of two opposite, simultaneous effects which have been modeled mathematically by fitting two parameter Weibull curves on the measured datas. Effects of the cylinder size and its geometrical relation on the measured strength values have been analyzed by finite element method. It was concluded that the geometry has a direct influence on the stress formation. Based on the results achieved, recommendations were given on how to perform the novel single fiber cylinder test

CurlySMILES: a chemical language to customize and annotate encodings of molecular and nanodevice structures

CurlySMILES is a chemical line notation which extends SMILES with annotations for storage, retrieval and modeling of interlinked, coordinated, assembled and adsorbed molecules in supramolecular structures and nanodevices. Annotations are enclosed in curly braces and anchored to an atomic node or at the end of the molecular graph depending on the annotation type. CurlySMILES includes predefined annotations for stereogenicity, electron delocalization charges, extra-molecular interactions and connectivity, surface attachment, solutions, and crystal structures and allows extensions for domain-specific annotations. CurlySMILES provides a shorthand format to encode molecules with repetitive substructural parts or motifs such as monomer units in macromolecules and amino acids in peptide chains. CurlySMILES further accommodates special formats for non-molecular materials that are commonly denoted by composition of atoms or substructures rather than complete atom connectivity

Identification and characterisation of enteroaggregative Escherichia coli subtypes associated with human disease

Enteroaggregative E. coli (EAEC) are a major cause of diarrhoea worldwide. Due to their heterogeneity and carriage in healthy individuals, identification of diagnostic virulence markers for pathogenic strains has been difficult. In this study, we have determined phenotypic and genotypic differences between EAEC strains of sequence types (STs) epidemiologically associated with asymptomatic carriage (ST31) and diarrhoeal disease (ST40). ST40 strains demonstrated significantly enhanced intestinal adherence, biofilm formation, and pro-inflammatory interleukin-8 secretion compared with ST31 isolates. This was independent of whether strains were derived from diarrhoea patients or healthy controls. Whole genome sequencing revealed differences in putative virulence genes encoding aggregative adherence fimbriae, E. coli common pilus, flagellin and EAEC heat-stable enterotoxin 1. Our results indicate that ST40 strains have a higher intrinsic potential of human pathogenesis due to a specific combination of virulence-related factors which promote host cell colonization and inflammation. These findings may contribute to the development of genotypic and/or phenotypic markers for EAEC strains of high virulence

Solution of the Schr\"odinger Equation for Quantum Dot Lattices with Coulomb Interaction between the Dots

The Schr\"odinger equation for quantum dot lattices with non-cubic, non-Bravais lattices built up from elliptical dots is investigated. The Coulomb interaction between the dots is considered in dipole approximation. Then only the center of mass (c.m.) coordinates of different dots couple with each other. This c.m. subsystem can be solved exactly and provides magneto- phonon like collective excitations. The inter-dot interaction is involved only through a single interaction parameter. The relative coordinates of individual dots form decoupled subsystems giving rise to intra-dot excitations. As an example, the latter are calculated exactly for two-electron dots. Emphasis is layed on qualitative effects like: i) Influence of the magnetic field on the lattice instability due to inter-dot interaction, ii) Closing of the gap between the lower and the upper c.m. mode at B=0 for elliptical dots due to dot interaction, and iii) Kinks in the single dot excitation energies (versus magnetic field) due to change of ground state angular momentum. It is shown that for obtaining striking qualitative effects one should go beyond simple cubic lattices with spherical dots. We also prove a more general version of the Kohn Theorem for quantum dot lattices. It is shown that for observing effects of electron- electron interaction between the dots in FIR spectra (breaking Kohn's Theorem) one has to consider dot lattices with at least two dot species with different confinement tensors.Comment: 11 figures included as ps-file