Quantification of the effects of antibodies on the extra- and intracellular dynamics of Salmonella enterica.

Antibodies are known to be essential in controlling Salmonella infection, but their exact role remains elusive. We recently developed an in vitro model to investigate the relative efficiency of four different human immunoglobulin G (IgG) subclasses in modulating the interaction of the bacteria with human phagocytes. Our results indicated that different IgG subclasses affect the efficacy of Salmonella uptake by human phagocytes. In this study, we aim to quantify the effects of IgG on intracellular dynamics of infection by combining distributions of bacterial numbers per phagocyte observed by fluorescence microscopy with a mathematical model that simulates the in vitro dynamics. We then use maximum likelihood to estimate the model parameters and compare them across IgG subclasses. The analysis reveals heterogeneity in the division rates of the bacteria, strongly suggesting that a subpopulation of intracellular Salmonella, while visible under the microscope, is not dividing. Clear differences in the observed distributions among the four IgG subclasses are best explained by variations in phagocytosis and intracellular dynamics. We propose and compare potential factors affecting the replication and death of bacteria within phagocytes, and we discuss these results in the light of recent findings on dormancy of Salmonella.

Closely correlating lower and upper bound plastic analysis of real slabs

An engineer assessing the load-carrying capacity of an existing reinforced concrete slab is likely to use elastic analysis to check the load at which the structure might be expected to fail in flexure or in shear. In practice, many reinforced concrete slabs are highly ductile in flexure, so an elastic analysis greatly underestimates the loads at which they fail in this mode. The use of conservative elastic analysis has led engineers to incorrectly condemn many slabs and therefore to specify unnecessary and wasteful flexural strengthening or replacement. The lower bound theorem is based on the same principles as the upper bound theorem used in yield line analysis, but any solution that rigorously satisfies the lower bound theorem is guaranteed to be a safe underestimate of the collapse load. Jackson presented a rigorous lower bound method that obtains very accurate results for complex real slabs.

Plastic limit solution for the response of plate anchors under combined translational and torsional undrained loading

Plate anchors are increasingly being used to moor large floating offshore structures in deep and ultradeep water. These facilities impart substantial vertical uplift loading to plate anchors. However, extreme operating conditions such as hurricane loading often result in partial system failures, with significant change in the orientation of the remaining intact mooring lines. The purpose of this study is to investigate the undrained pure translational (parallel to plate) and torsional bearing capacity of anchor plates idealized as square and rectangular shaped plates. Moreover, the interaction response of plate anchors under combined translational and torsional loading is studied using a modified plastic limit analysis (PLA) approach. The previous PLA formulation which did not account for shear-normal force interaction on the vertical end faces of the plate provides an exact solution to the idealized problem of an infinitely thin plate but only an approximate solution to the problem of a plate of finite thickness. This is also confirmed by the three-dimensional finite element (FE) results, since the PLA values exceed FE results as the thickness of the plate increases. By incorporating the shear-normal interaction relationship in the modified solution, the torsional bearing capacity factors, as well as the plate interaction responses are enhanced as they show satisfactory agreement with the FE results. The interaction relationship is then obtained for square and rectangular plates of different aspect ratios and thicknesses. The new interaction relationships could also be used as an associated plastic failure locus for combined shear and torsional loading to predict plastic displacements and rotations in translational and torsional loading modes as well. Copyright © 2011 by the International Society of Offshore and Polar Engineers (ISOPE).

3 Gbit/s LED-based step index plastic optical fiber link using multilevel pulse amplitude modulation

Multilevel PAM is investigated for a LED-based SI-POF link. Using PAM-8, transmission at a record 3 Gbit/s is demonstrated for a maximum length of 25 m step index POF with offline post-receiver processing. © 2013 OSA.

FEC-free 50 m 1.5 Gb/s plastic optical fibre link using CAP modulation for home networks

LED-based carrierless amplitude and phase modulation is investigated for a multi-gigabit plastic optical fibre link. An FPGA-based 1.5 Gbit/s error free transmission over 50 m standard SI-POF using CAP64 is achieved, providing 2.9 dB power margin without forward error correction. © 2012 OSA.

3 Gbit/s LED-based step index plastic optical fiber link using multilevel pulse amplitude modulation

Multilevel PAM is investigated for a LED-based SI-POF link. Using PAM-8, transmission at a record 3 Gbit/s is demonstrated for a maximum length of 25 m step index POF with offline post-receiver processing. © 2013 OSA.

Optimal displacement mechanisms beneath shallow foundations on linear-elastic perfectly plastic soil

An energy method for a linear-elastic perfectly plastic method utilising the von Mises yield criterion with associated flow developed in 2013 by McMahon and co-workers is used to compare the ellipsoidal cavity-expansion mechanism, from the same work, and the displacement fields of other research by Levin, in 1995, and Osman and Bolton, in 2005, which utilise the Hill and Prandtl mechanisms respectively. The energy method was also used with a mechanism produced by performing a linear-elastic finite-element analysis in Abaqus. At small values of settlement and soil rigidity the elastic mechanism provides the lowest upper-bound solution, and matches well with finite-element analysis results published in the literature. At typical footing working loads and settlements the cavity-expansion mechanism produces a more optimal solution than the displacement fields within the Hill and Prandtl mechanisms, and also matches well with the published finite-element analysis results in this range. Beyond these loads, at greater footing settlements, or soil rigidity, the Prandtl mechanism is shown to be the most appropriate.

FEC-free 50 m 1.5 Gb/s plastic optical fibre link using CAP modulation for home networks

LED-based carrierless amplitude and phase modulation is investigated for a multi-gigabit plastic optical fibre link. An FPGA-based 1.5 Gbit/s error free transmission over 50 m standard SI-POF using CAP64 is achieved, providing 2.9 dB power margin without forward error correction. © 2012 Optical Society of America.

Detection of cutaneous antibodies in excised skin explants from grass carp, Ctenopharyngodon idella (Valenciennes), immune to Scophthalmus maximus rhabdovirus

This study determined whether cutaneous antibodies were present in excised skin explants of grass carp, Ctenopharyngodon idella, immune to Scophthalmus maximus rhabdovirus (SMRV). Culture fluid from immune skin explants were assayed by indirect enzyme-linked immunosorbent assay (iELISA), Western blot, indirect immunofluorescent assay (IFA) and flow cytometry (FCM). iELISA showed that cutaneous antibody titres were much lower (1:12) than antiserum titres (1:1458) from intraperitoneally immunized grass carp. The phosphoprotein and matrix protein antigens of purified SMRV proteins were recognized by cutaneous antibodies from skin culture fluid using Western blot. The skin culture fluid produced staining signals in viral assembly sites and cytoplasm of SMRV-infected epithelioma papulosum cyprini (EPC) cells by IFA. FCM showed that 4.39% SMRV-infected EPC cells were detected, while non-specific reaction was seen in 2% of control cells. This is the first description of cutaneous antibodies against SMRV in grass carp.

Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with-full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron Microscopy, Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV. (c) 2008 Published by Elsevier Inc.

Development and characterization of monoclonal antibodies to spring viraemia of carp virus

Five monoclonal antibodies (mAbs) against spring viraemia of carp (SVCV0504, isolated from common carp in China) were produced from mice immunized with purified virus preparations. The virion of SVCV contains five structural proteins, representing the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (Q. Western blotting analysis revealed that three mAbs (1145, IE10, and 11-17) recognized specifically to a single protein of 47 kDa (N), the mAb 3G4 reacted with, two SVCV0504 proteins of 69 kDa (G) and 47 kDa (N), while the mAb 1A9 reacted with three SVCV0504 proteins of 69 kDa (G), 50 kDa (P), and 47 kDa (N). By indirect ELISA, two mAbs (1H5 and 11-17) showed cross-reactivity with pike fry rhabdovirus (PFRV), but no cross-reactions with the Siniperca chuatsi rhabdovirus (SCRV), Scophthalmus maximus rhabdovirus (SMRV), Paralichthys olivaceus rhabdovirus (PoRV) were demonstrated with the five mAbs. Indirect immunofluorescence showed intense fluorescence in the cytoplasm of the SVCV0504-infected epithelioma papulosum cyprini (EPC) cells in areas corresponding to the location of granular structures. The sucrose gradient-purified SVCV0504 particles could be detected successfully by these mAbs using immunodot blotting. mAb 1A9 could completely neutralize 100 TCID50 (50% tissue culture infective dose) of SVCV0504 at a dilution of 1:8. This is the first report of development of the neutralizing mAbs against SVCV. The mAb 1A9 was analyzed further and could be used to successfully detect viral antigens in the infected-EPC cell cultures or in cryosections from experimentally infected crucian carp (Carassius auratus) by immunohistochemistry assay. Furthermore, a flow cytometry procedure for the detection and quantification of cytoplasmic SVCV0504 in cell cultures was developed with mAb 1A9. At 28 h after inoculation with the virus (0.01 PFU/cell), 10.12% of infected cells could be distinguished from the uninfected cells. These mAbs will be useful in diagnostic test development and pathogenesis studies for fish rhabdovirus. (c) 2008 Elsevier B.V. All rights reserved.

Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus

Two MAbs (3C7 and 3C9) against flounder Paralichthys olivaceus rhabdovirus (PORV) were generated with hybridoma cell fusion technology and characterized by an indirect enzyme-linked immunosorbent assay, isotype test, Western blot and immunodot analysis and immunofluorescence assay. Isotyping tests demonstrated that both of the two MAbs belonged to IgM subclass. Western blot analysis showed the MAbs reacted with 42, 30, and 22 kDa viral proteins, which were localized within the cytoplasm of PORV-infected grass carp ovary (GCO) cells analyzed by indirect immunofluorescences tests. The MAb 3C7 was also selected at random for detecting virus antigens in the inoculated grass carp tissues by immunohistochemistry assay. Flow cytometry tests showed that at the 36 h postinfection (0.25 PFU/cell), the 23% PORV-infected GCO cells could be distinguished from the uninfected cells with the MAb 3C7. Such MAbs could be useful for diagnosis and potential treatment of viral infection. (C) 2007 Elsevier B.V. All rights reserved.