Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules

<p>Abstract</p> <p>Background</p> <p>Protein A, protein G and protein L are three well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Although the precise functions of these molecules are not fully understood, it is thought that they play an important role in pathogenicity of bacteria. The single domains of protein A, protein G and protein L were all demonstrated to have function to bind to Ig. Whether combinations of Ig-binding domains of various IBPs could exhibit useful novel binding is interesting.</p> <p>Results</p> <p>We used a combinatorial phage library which displayed randomly-rearranged various-peptide-linked molecules of D and A domains of protein A, designated PA(D) and PA(A) respectively, B2 domain of protein G (PG) and B3 domain of protein L (PL) for affinity selection with human IgG (hIgG), human IgM (hIgM), human IgA (hIgA) and recombinant hIgG1-Fc as bait respectively. Two kinds of novel combinatorial molecules with characteristic structure of PA(A)-PG and PA(A)-PL were obtained in hIgG (hIgG1-Fc) and hIgM (hIgA) post-selection populations respectively. In addition, the linking peptides among all PA(A)-PG and PA(A)-PL structures was strongly selected, and showed interestingly divergent and convergent distribution. The phage binding assays and competitive inhibition experiments demonstrated that PA(A)-PG and PA(A)-PL combinations possess comparable binding advantages with hIgG/hIgG1-Fc and hIgM/hIgA respectively.</p> <p>Conclusion</p> <p>In this work, a combinatorial phage library displaying Ig-binding domains of protein A, protein G, or protein L joined by various random linking peptides was used to conducted evolutional selection <it>in vitro</it> with four kinds of Ig molecules. Two kinds of novel combinations of Ig-binding domains, PA(A)-PG and PA(A)-PL, were obtained, and demonstrate the novel Ig binding properties.</p

Colouring (Pr+Ps)-free graphs.

The k-Colouring problem is to decide if the vertices of a graph can be coloured with at most k colours for a fixed integer k such that no two adjacent vertices are coloured alike. If each vertex u must be assigned a colour from a prescribed list L(u) subseteq {1,...,k}, then we obtain the List k-Colouring problem. A graph G is H-free if G does not contain H as an induced subgraph. We continue an extensive study into the complexity of these two problems for H-free graphs. We prove that List 3-Colouring is polynomial-time solvable for (P_2+P_5)-free graphs and for (P_3+P_4)-free graphs. Combining our results with known results yields complete complexity classifications of 3-Colouring and List 3-Colouring on H-free graphs for all graphs H up to seven vertices. We also prove that 5-Colouring is NP-complete for (P_3+P_5)-free graphs

A neural network model for constructing endophenotypes of common complex diseases: an application to male young-onset hypertension microarray data

Motivation: Identification of disease-related genes using high-throughput microarray data is more difficult for complex diseases as compared with monogenic ones. We hypothesized that an endophenotype derived from transcriptional data is associated with a set of genes corresponding to a pathway cluster. We assumed that a complex disease is associated with multiple endophenotypes and can be induced by their up/downregulated gene expression patterns. Thus, a neural network model was adopted to simulate the gene–endophenotype–disease relationship in which endophenotypes were represented by hidden nodes

DEM numerical investigation of wet particle flow behaviors in multiple-spout fluidized beds

Spout fluidized beds are important for industrial processing, and multiple-spout fluidized beds play an important role in chemical reactions. However, particle flow behaviors in multiple-spout fluidized beds are not well known in wet particle systems. In this study, the flow behaviors of particles were investigated in dry and humid multiple-spout fluidized beds using a discrete element method (DEM). The simulated spout fluidized beds are similar to the ones used in the Buijtenen et al.’s experiment (published in Chemical Engineering Science, 2011, 66(11): 2368-2376). In the reference, particle flow behaviors were measured and investigated by PIV and PEPT in multiple spout fluidized beds. In this work, the simulated results are compared with the experimental data in single and double spout fluidized beds from Buijtenen et al., and the time-averaged particle velocities are compared to validate the simulation method. In contrast, simulated results with a liquid content of 1% in the bed showed good agreement with the data in the experimental results with an air relative humidity of 50%. Different liquid contents of the particles were applied to investigate the particle flow behaviors in wet granular systems. The liquid bridge force had a strong influence on the flow behaviors of the particles in the dense region, which resulted in different hydrodynamic characteristics between the dry and wet particles. In addition, the drag force dominated the particle flow behavior in the dry and wet particle systems. Moreover, in a wet granular system, the mass particle fluxes decreased, and the fluctuation of the pressure drops increased with an increasing influence of the liquid bridge force on the particles. Furthermore, with an increasing liquid content, the energy fluctuation of the particles and bubbles weakened gradually with less active motions. A comparison of the hydrodynamic flow behaviors in single-spout and double-spout fluidized beds was carried out as well. Comparisons of the solid circulation rate and the colliding characteristics between single-spout and double-spout fluidized beds were conducted. Particularly, a comparison of the mixing characteristics demonstrated that the particles were mixed more completely in a double-spout fluidized bed. Therefore, the double-spout fluidized bed could provide more adequate space for mass and heat transfer under the same condition. This was important in providing a theory for designing the industrial reactor

Epstein–Barr Virus DNase (BGLF5) induces genomic instability in human epithelial cells

Epstein–Barr Virus (EBV) DNase (BGLF5) is an alkaline nuclease and has been suggested to be important in the viral life cycle. However, its effect on host cells remains unknown. Serological and histopathological studies implied that EBV DNase seems to be correlated with carcinogenesis. Therefore, we investigate the effect of EBV DNase on epithelial cells. Here, we report that expression of EBV DNase induces increased formation of micronucleus, an indicator of genomic instability, in human epithelial cells. We also demonstrate, using γH2AX formation and comet assay, that EBV DNase induces DNA damage. Furthermore, using host cell reactivation assay, we find that EBV DNase expression repressed damaged DNA repair in various epithelial cells. Western blot and quantitative PCR analyses reveal that expression of repair-related genes is reduced significantly in cells expressing EBV DNase. Host shut-off mutants eliminate shut-off expression of repair genes and repress damaged DNA repair, suggesting that shut-off function of BGLF5 contributes to repression of DNA repair. In addition, EBV DNase caused chromosomal aberrations and increased the microsatellite instability (MSI) and frequency of genetic mutation in human epithelial cells. Together, we propose that EBV DNase induces genomic instability in epithelial cells, which may be through induction of DNA damage and also repression of DNA repair, subsequently increases MSI and genetic mutations, and may contribute consequently to the carcinogenesis of human epithelial cells

Loss of Cofilin 1 Disturbs Actin Dynamics, Adhesion between Enveloping and Deep Cell Layers and Cell Movements during Gastrulation in Zebrafish

During gastrulation, cohesive migration drives associated cell layers to the completion of epiboly in zebrafish. The association of different layers relies on E-cadherin based cellular junctions, whose stability can be affected by actin turnover. Here, we examined the effect of malfunctioning actin turnover on the epibolic movement by knocking down an actin depolymerizing factor, cofilin 1, using antisense morpholino oligos (MO). Knockdown of cfl1 interfered with epibolic movement of deep cell layer (DEL) but not in the enveloping layer (EVL) and the defect could be specifically rescued by overexpression of cfl1. It appeared that the uncoordinated movements of DEL and EVL were regulated by the differential expression of cfl1 in the DEL, but not EVL as shown by in situ hybridization. The dissociation of DEL and EVL was further evident by the loss of adhesion between layers by using transmission electronic and confocal microscopy analyses. cfl1 morphants also exhibited abnormal convergent extension, cellular migration and actin filaments, but not involution of hypoblast. The cfl1 MO-induced cell migration defect was found to be cell-autonomous in cell transplantation assays. These results suggest that proper actin turnover mediated by Cfl1 is essential for adhesion between DEL and EVL and cell movements during gastrulation in zebrafish