Human Intestinal Cells Modulate Conjugational Transfer of Multidrug Resistance Plasmids between Clinical Escherichia coli Isolates.

Bacterial conjugation in the human gut microbiota is believed to play a major role in the dissemination of antibiotic resistance genes and virulence plasmids. However, the modulation of bacterial conjugation by the human host remains poorly understood and there is a need for controlled systems to study this process. We established an in vitro co-culture system to study the interaction between human intestinal cells and bacteria. We show that the conjugation efficiency of a plasmid encoding an extended spectrum beta-lactamase is reduced when clinical isolates of Escherichia coli are co-cultured with human intestinal cells. We show that filtered media from co-cultures contain a factor that reduces conjugation efficiency. Protease treatment of the filtered media eliminates this inhibition of conjugation. This data suggests that a peptide or protein based factor is secreted on the apical side of the intestinal cells exposed to bacteria leading to a two-fold reduction in conjugation efficiency. These results show that human gut epithelial cells can modulate bacterial conjugation and may have relevance to gene exchange in the gut

Effect of chain flexibility in the network structure on the viscoelasticity of epoxy thermosets

In this paper a detailed study in investigating the effect of the chain flexibility in epoxy-amine crosslinked network is done. In order to introduce flexibility into the crosslinked network a homologous series of four aliphatic diamine curing agents varying only in the chain length and having a constant functionality (f?=?4) is taken and cured stoichiometrically with aromatic epoxy (f?=?2) resin. For each of the cured mixture the viscoelastic master curve and corresponding shift factors were determined. It is found the introduction of flexibility shifts the viscoelastic curves by 5 decades with respect to frequency scale. This shift in the viscoelastic curve is modeled with a parameterized Havriliak-Negami model for the master curve. The free volume contribution for the changes in the coefficient of thermal expansion at T g is also determined.Precision and Microsystems EngineeringMechanical, Maritime and Materials Engineerin

Viral Metagenomics Approaches for High-Resolution Screening of Multiplexed Arthropod and Plant Viral Communities

Viral metagenomic approaches have become essential for culture-independent and sequence-independent viral detection and characterization. This chapter describes an accurate and efficient approach to (1) concentrate viral particles from arthropods and plants, (2) remove contaminating non-encapsidated nucleic acids, (3) extract and amplify both viral DNA and RNA, and (4) analyze high-throughput sequencing (HTS) data by bioinformatics. Using this approach, up to 96 arthropod or plant samples can be multiplexed in a single HTS library