Caf1A usher possesses a Caf1 subunit-like domain that is crucial for Caf1 fibre secretion

The chaperone/usher pathway controls assembly of fibres of adhesive organelles of Gram-negative bacteria. The final steps of fibre assembly and fibre translocation to the cell surface are co-ordinated by the outer membrane proteins, ushers. Ushers consist of several soluble periplasmic domains and a single transmembrane beta-barrel. Here we report isolation and structural/functional characterization of a novel middle domain of the Caf1A usher from Yersinia pestis. The isolated UMD (usher middle domain) is a highly soluble monomeric protein capable of autonomous folding. A 2.8 angstrom (1 angstrom = 0.1 nm) resolution crystal structure of UMD revealed that this domain has an immunoglobulin-like fold similar to that of donor-strand-complemented Caf1 fibre subunit. Moreover, these proteins displayed significant structural similarity. Although UMD is in the middle of the predicted amphipathic beta-barrel of Caf1A, the usher still assembled in the membrane in the absence of this domain. UMD did not bind Caf1M-Caf1 complexes, but its presence was shown to be essential for Caf1 fibre secretion. The study suggests that UMD may play the role of a subunit-substituting protein (dummy subunit), plugging or priming secretion through the channel in the Caf1A usher. Comparison of isolated UMD with the recent strcture of the corresponding domain of PapC usher revealed high similarity of the core structures, suggesting a universal structural adaptation of FGL (F(1)G(1) long) and FGS (F(1)G(1) short) chaperone/usher pathways for the secretion of different types of fibres. The functional role of two topologically different states of this plug domain suggested by structural and biochemical results is discussed.

Resolving the energy paradox of chaperone/usher-mediated fibre assembly

Periplasmic chaperone/usher machineries are used for assembly of filamentous adhesion organelles of Gram-negative pathogens in a process that has been suggested to be driven by folding energy. Structures of mutant chaperone-subunit complexes revealed a final folding transition (condensation of the subunit hydrophobic core) on the release of organelle subunit from the chaperone-subunit pre-assembly complex and incorporation into the final fibre structure. However, in view of the large interface between chaperone and subunit in the pre-assembly complex and the reported stability of this complex, it is difficult to understand how final folding could release sufficient energy to drive assembly. In the present paper, we show the X-ray structure for a native chaperone-fibre complex that, together with thermodynamic data, shows that the final folding step is indeed an essential component of the assembly process. We show that completion of the hydrophobic core and incorporation into the fibre results in an exceptionally stable module, whereas the chaperone-subunit preassembly complex is greatly destabilized by the high-energy conformation of the bound subunit. This difference in stabilities creates a free energy potential that drives fibre formation.

Modelling the cabling of rope systems

This paper presents a theoretical model of the torsional characteristics of parallel multi-part rope systems. In such systems, the ropes may cable, or wrap around each other, depending on the combination of applied torque, rope tension, length and spacing between the rope parts. Cabling constitutes a failure that might be retrievable but as such can seriously affect the performance of the rope system. The torsional characteristics of the system are very different before and after cabling, and theoretical models are given for both situations. Laboratory tests were performed on both two and four rope systems, with measurements being made of torque at rotations from 0 to 360 deg. Tests were run with different rope spacings, tensions and lengths and the results compared with predictions from the theoretical model. The conclusion from the test results was that the theoretical model predicts both the pre- and post-cabling torsional behaviour with an acceptable level of accuracy.

Design, testing, and simulation of fibre composite leaf springs for heavy axle loads

This paper summarizes the design, manufacturing, testing, and finite element analysis (FEA) of glass-fibre-reinforced polyester leaf springs for rail freight vehicles. FEA predictions of load-deflection curves under static loading are presented, together with comparisons with test results. Bending stress distribution at typical load conditions is plotted for the springs. The springs have been mounted on a real wagon and drop tests at tare and full load have been carried out on a purpose-built shaker rig. The transient response of the springs from tests and FEA is presented and discussed.

Use of thermography and fibre optics microscopy for the evaluation of railway structures in the Athens Metro in Greece (Invited Paper)

Rolling Contact Fatigue (RCF) is one of the main issues that concern, at least initially, the head of the railway; progressively they can be of very high importance as they can propagate inside the material with the risk of damaging the railway. In this work, two different non-destructive techniques, infrared thermography (IRT) and fibre optics microscopy (FOM), were used in the inspection of railways for the tracing of defects and deterioration signs. In the first instance, two different approaches (dynamic and pulsed thermography) were used, whilst in the case of FOM, microscopic characterisation of the railway heads and classification of the deterioration -- damage on the railways according to the UIC (International Union of Railways) code, took place. Results from both techniques are presented and discussed.

Impact properties of glass/plant fibre hybrid laminates

The use of plants fibre reinforced composites has continuously increased during recent years. Their low density, higher environmental friendliness, and reduced cost proved particularly attractive for low-tech applications e.g., in building, automotive and leisure time industry. However, a major limitation to the use of these materials in structural components is unsatisfactory impact performance. An intermediate approach, the production of glass/ plant fibre hybrid laminates, has also been explored, trying to obtain materials with sufficient impact properties, whilst retaining a reduced cost and a substantial environmental gain. A survey is given on some aspects, crucial for the use of glass/plant fibre hybrid laminates in structural components: performance of hybrids when subjected to impact testing; the effect of laminate configuration, manufacturing procedure and fibre treatment on impact properties of the composite. Finally, indications are provided for a suitable selection of plant fibres with minimal extraction damage and sufficient toughness, for introduction in an impact-resistant glass/plant fibre hybrid laminate.

Mapping fibre orientation in complex-shaped biological systems with micrometre resolution by scanning X-ray microdiffraction

A fully automated procedure to extract and to image local fibre orientation in biological tissues from scanning X-ray diffraction is presented. The preferred chitin fibre orientation in the flow sensing system of crickets is determined with high spatial resolution by applying synchrotron radiation based X-ray microbeam diffraction in conjunction with advanced sample sectioning using a UV micro-laser. The data analysis is based on an automated detection of azimuthal diffraction maxima after 2D convolution filtering (smoothing) of the 2D diffraction patterns. Under the assumption of crystallographic fibre symmetry around the morphological fibre axis, the evaluation method allows mapping the three-dimensional orientation of the fibre axes in space. The resulting two-dimensional maps of the local fibre orientations - together with the complex shape of the flow sensing system - may be useful for a better understanding of the mechanical optimization of such tissues.

Potential prebiotic activity of oligosaccharides obtained by enzymatic conversion of durum wheat insoluble dietary fibre into soluble dietary fibre

Background and aims: Epidemiological evidence indicates that cereal dietary fibre (DF) may have several cardiovascular health benefits. The underlying mechanisms have not yet been elucidated. Here, the potential nutritional effects of physico-chemical. properties modifications of durum wheat dietary fibre (DWF) induced by enzyme treatment have been investigated. Methods and results: The conversion of the highly polymerised insoluble dietary fibre into soluble feruloyl oligosaccharides of DWF was achieved by a tailored enzymatic treatment. The in vitro fermentation and release of ferulic acid by intestinal microbiota from DWF before and after the enzymatic treatment were assessed using a gut model validated to mimic the human colonic microbial environment. Results demonstrated that, compared to DWF, the enzyme-treated DWF (ETD-WF) stimulated the growth of bifidobacteria and lactobacilli. Concurrently, the release of free ferulic acid by ET-DWF was almost three times higher respect to the control. No effect on the formation of short chain fatty acids was observed. Conclusions: The conversion of insoluble dietary fibre from cereals into soluble dietary fibre generated a gut microbial fermentation that supported bifidobacteria and lactobacilli. The concurrent increase in free ferulic acid from the enzyme-treated DWF might result in a higher plasma ferulic acid concentration which could be one of the reasons for the health benefits reported for dietary fibre in cardiovascular diseases. (c) 2008 Elsevier B.V. All rights reserved.

Fibre and effects on probiotics (the prebiotic concept)

The burden (economic and medicinal) of acute and chronic gut disorders continues to increase. As efficient therapies are few, attention has turned towards the use of so-called functional foods to mediate against gut disorder. These target particular genera of gut bacteria seen as beneficial, e.g. bifidobacteria, lactobacilli. The use of products containing live microbial species (probiotics) has a long history of use in humans and many trials have been reported as 'positive'. Taking the view that positive components of the gut flora already exist in the intestinal tract, the prebiotic concept has been developed. Here, dietary carbohydrates have a selective metabolism within the gut flora thereby shifting the community towards a more advantageous structure. Conventional fibres like pectins, cellulose, etc. are not selectively metabolised by gut bacteria. However, certain oligosaccharides do have this capability. Most research has been conducted with fructooligosaccharides, like inulin, which have a powerful bifidogenic effect. Trials are ongoing to determine the clinical benefits of prebiotic use. Intestinal disorders like ulcerative colitis, gastroenteritis and irritable bowel syndrome are particular targets. (c) 2004 Elsevier Ltd. All rights reserved.