Pasteurella multocida Expresses Two Lipopolysaccharide Glycoforms Simultaneously, but Only a Single Form Is Required for Virulence: Identification of Two Acceptor-Specific Heptosyl I Transferases

Lipopolysaccharide (LPS) is a critical virulence determinant in Pasteurella multocida and a major antigen responsible for host protective immunity. In other mucosal pathogens, variation in LPS or lipooligosaccharide structure typically occurs in the outer core oligosaccharide regions due to phase variation. P. multocida elaborates a conserved oligosaccharide extension attached to two different, simultaneously expressed inner core structures, one containing a single phosphorylated 3-deoxy-D-manno-octulosonic acid (Kdo) residue and the other containing two Kdo residues. We demonstrate that two heptosyltransferases, HptA and HptB, add the first heptose molecule to the Kdo1 residue and that each exclusively recognizes different acceptor molecules. HptA is specific for the glycoform containing a single, phosphorylated Kdo residue (glycoform A), while HptB is specific for the glycoform containing two Kdo residues (glycoform B). In addition, KdkA was identified as a Kdo kinase, required for phosphorylation of the first Kdo molecule. Importantly, virulence data obtained from infected chickens showed that while wild-type P. multocida expresses both LPS glycoforms in vivo, bacterial mutants that produced only glycoform B were fully virulent, demonstrating for the first time that expression of a single LPS form is sufficient for P. multocida survival in vivo. We conclude that the ability of P. multocida to elaborate alternative inner core LPS structures is due to the simultaneous expression of two different heptosyltransferases that add the first heptose residue to the nascent LPS molecule and to the expression of both a bifunctional Kdo transferase and a Kdo kinase, which results in the initial assembly of two inner core structures.

Thin-Walled Timber Structures

Due to their efficiency, lightweight, ease of erection and low cost, steel and aluminium thin-walled structures have become very popular in the construction industry over the past few decades. Applications include roof and wall systems (purlins and girts), storage racks, and composite concrete and steel slabs. The effectiveness of these structures lies in the cross-sectional shape of the profiles which enhances their strength by controlling the three fundamental buckling modes: local, distortional, and global. However, despite the attractiveness of these structures, steel and aluminium are greenhouse gas intensive materials and do not produce sustainable structural products. This paper presents an investigation performed at the Griffith School of Engineering, Griffith University, which shows manufacturing these types of profiles in timber is possible. Short composite thinwalled timber Cee-sections (500 mm long) were fabricated by gluing together thin softwood (Araucaria cunninghamii) veneers (1 mm thick). Two types of Ceesections were considered, one with a web stiffener to increase the local buckling capacity of the profile and one without. The profiles were tested in compression and the test results are presented and discussed in the paper in terms of structural behaviour and performance. Further research directions are proposed in order to provide efficient and lightweight sustainable structural products to the timber industry. © RILEM 2014.

Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures

Using benzene as a candidate air toxicant and A549 cells as an in vitro cell model, we have developed and validated a hanging drop (HD) air exposure system that mimics an air liquid interface exposure to the lung for periods of 1 h to over 20 days. Dose response curves were highly reproducible for 2D cultures but more variable for 3D cultures. By comparing the HD exposure method with other classically used air exposure systems, we found that the HD exposure method is more sensitive, more reliable and cheaper to run than medium diffusion methods and the CULTEX (R) system. The concentration causing 50% of reduction of cell viability (EC50) for benzene, toluene, p-xylene, m-xylene and o-xylene to A549 cells for 1 h exposure in the HD system were similar to previous in vitro static air exposure. Not only cell viability could be assessed but also sub lethal biological endpoints such as DNA damage and interleukin expressions. An advantage of the HD exposure system is that bioavailability and cell concentrations can be derived from published physicochemical properties using a four compartment mass balance model. The modelled cellular effect concentrations EC50(cell) for 1 h exposure were very similar for benzene, toluene and three xylenes and ranged from 5 to 15 mmol/kg(dry weight) which corresponds to the intracellular concentration of narcotic chemicals in many aquatic species, confirming the high sensitivity of this exposure method. (C) 2013 Elsevier B.V. All rights reserved.

Thin-walled timber and FRP-timber veneer composite CEE-sections

This paper compares the structural performance between thin-walled timber and FRP-timber composite Cee-sections. While, thin-walled composite timber structures have been proven to be efficient and ultra-light structural elements, their manufacturing is difficult and labour intensive. Significant effort and time is required to prevent the cracking of the transverse timber veneers, bent in the grain direction, when forming the cross-sectional shape. FRP-timber structures overcome this disadvantage by replacing the transverse veneers with flexible, unidirectional FRP material and only keeping the timber veneers which are bent in their natural rolling direction. The Cee-sections investigated in this study were 210 mm deep × 90 mm wide × 500 mm high and manufactured from five plies. For both section types, the three internal plies were thin (1 mm thick) softwood Hoop pine (Araucaria cunninghamii) veneers, orientated along the section longitudinal axis. The two outer layers, providing bending stiffness to the walls, were Hoop pine veneers (1 mm thick) for the timber sections and glass fibre reinforced plastic (0.73 mm thick) for the FRP-timber sections orientated perpendicular to the inner layers. The manufacturing process is briefly introduced in this paper. The profiles were fitted with strain gauges and tested in compression. Linear Variable Displacement Transducers also recorded the buckling along one flange. The test results are presented and discussed in this paper in regards to their structural behaviour and performance. Results showed that the use of FRP in the sections increases both the elastic local buckling load and section capacity, the latter being increased by about 24 percent. The results indicate that thin-walled FRP-timber can ultimately be used as a sustainable alternative to cold-formed steel profiles.

Three-dimensional reconstruction of black tiger prawn (Penaeus monodon) spermatozoa using serial block-face scanning electron microscopy

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) was used in this study to examine the ultrastructural morphology of Penaeus monodon spermatozoa. SBF-SEM provided a large dataset of sequential electron-microscopic-level images that facilitated comprehensive ultrastructural observations and three-dimensional reconstructions of the sperm cell. Reconstruction divulged a nuclear region of the spermatophoral spermatozoon filled with decondensed chromatin but with two apparent levels of packaging density. In addition, the nuclear region contained, not only numerous filamentous chromatin elements with dense microregions, but also large centrally gathered granular masses. Analysis of the sperm cytoplasm revealed the presence of degenerated mitochondria and membrane-less dense granules. A large electron-lucent vesicle and "arch-like" structures were apparent in the subacrosomal area, and an acrosomal core was found in the acrosomal vesicle. The spermatozoal spike arose from the inner membrane of the acrosomal vesicle, which was slightly bulbous in the middle region of the acrosomal vesicle, but then extended distally into a broad dense plate and to a sharp point proximally. This study has demonstrated that SBF-SEM is a powerful technique for the 3D ultrastructural reconstruction of prawn spermatozoa, that will no doubt be informative for further studies of sperm assessment, reproductive pathology and the spermiocladistics of penaeid prawns, and other decapod crustaceans. J. Morphol., 2016. (c) 2016 Wiley Periodicals, Inc.