Search Results (15)

In this modern era, the constant increase in computational and sensing power has lead to the development of multiple data-driven concepts. Amongst these, the ‘digital thread’ is an architecture that aims at optimising the knowledge of a system by merging prior knowledge of the product with information from multiple stages of its lifecycle in order to improve the performance of new products to be designed, thanks to the increased accuracy of this updated knowledge. Even though the use of these data-driven architectures is becoming increasingly widespread, most of the corresponding developments remain currently limited to the component level. In this respect, this article extends the application of the digital thread from the component level to the structural assembly level and enriches it with additional multi-physics considerations and non-linear failure constraints. To this end, it details the development of a digital thread dedicated to an aircraft vertical tail plane structure made of carbon fibre reinforced polymer, as well as the tools and models required to implement such an approach using Bayesian inference, multi-physics simulations, and empirical models. Full article

Cattle are increasingly exposed to hot temperatures as a result of climate change, and a better understanding of behavioural responses could be beneficial for the diagnosis of heat loads. The changes in the positioning of key body parts, feeding behaviour, body maintenance, and respiratory dynamics were assessed in 24 Black Angus steers individually exposed to hot conditions and fed a finisher diet based on cereal grain or a substituted diet (8% of the grain replaced by an isoenergetic amount of lucerne hay). Increased respiration rate during the heat load period, compared to the recovery period, was associated with increased stepping, especially by left limbs. Cattle also reduced eating, grooming, and scratching during the heat load period. The lowered head, backward ear, vertical or raised tail, and increased respiration rate and panting persisted in cattle during the heat load period. Cattle on the cereal grain diet stood for longer and were more likely to hold their ears backward and tail vertical than those on the substituted diet. We conclude that these behaviours could be used to detect animals that are most affected and that changing from a cereal-based diet to a substituted diet containing a higher amount of fibre, such as lucerne hay, can reduce hyperthermic behavioural responses to a heat load. Full article

This paper describes the transition towards a composite structure, with the same overall aerodynamic characteristics, for a tail rotor blade of an IAR330 helicopter. The newly proposed structure of the composite blade is made of a carbon-roving spar embedded with epoxy resin, a hexagonal-cell honeycomb core manufactured by fused deposition modelling, and an outer skin made of multiple carbon-fibre-reinforced laminae. The blade was manufactured by the authors using the hand lay-up method at a scale of 1:3 with respect to the real one, and all stages of the manufacturing process are extensively described in the paper. The experimental tests were performed on an Instron 8872 testing machine by applying a bending force on its free edge, similar to the testing methodology employed by various composite blade manufacturers. A three-dimensional numerical model of the tail rotor blade was conceived, analysed using the finite element method, and validated by comparing the numerical and experimental values of the maximum bending force. Further, the model was used for a complex finite element analysis that showed the very good behaviour of the proposed composite blade during flight and emphasized the main advantages brought by the proposed composite structure. Full article

Medicinal mushrooms, e.g., Lion’s Mane (Hericium erinaceus (Bull.) Pers.), Reishi (Ganoderma lucidum (Curtis) P. Karst.), Chaga (Inonotus obliquus (Ach. ex Pers.) Pilát), Cordyceps (Ophiocordyceps sinensis (Berk.) G.H. Sung, J.M. Sung, Hywel-Jones and Spatafora), Shiitake (Lentinula edodes (Berk.) Pegler), and Turkey Tail (Trametes versicolor (L.) Lloyd), are considered new-generation foods and are of growing interest to consumers. They are characterised by a high content of biologically active compounds, including (1,3)(1,6)-β-d-glucans, which are classified as dietary fibre, triterpenes, phenolic compounds, and sterols. Thanks to their low-fat content, they are a low-calorie product and are classified as a functional food. They have a beneficial effect on the organism through the improvement of its overall health and nutritional level. The biologically active constituents contained in medicinal mushrooms exhibit anticancer, antioxidant, antidiabetic, and immunomodulatory effects. In addition, these mushrooms accelerate metabolism, help fight obesity, and slow down the ageing processes thanks to their high antioxidant activity. The vast therapeutic properties of mushrooms are still not fully understood. Detailed mechanisms of the effects of medicinal mushrooms on the human organism still require long-term clinical studies to confirm their nutraceutical effects, their safety of use, and their dosage. Medicinal mushrooms have great potential to be used in the design of innovative functional foods. There is a need for further research on the possibility of incorporating mushrooms into food products to assess the interactions of their bioactive substances with ingredients in the food matrix. This review focuses on the properties of selected medicinal mushrooms and their effects on the human organism and presents current knowledge on the possibilities of their use in the production of functional foods. Full article

In order to solve the limitation of graphite tailings in improving the toughness of cement-based materials, this paper aims to study the effect of basalt fibres (BF) on the mechanical properties of graphite tailings cement mortar (GTCM). Basic physical and mechanical tests such as fluidity, water absorption, surface water content, flexural strength, compressive strength and modulus of elasticity were conducted on basalt fibre-reinforced graphite tailings cement mortar (BFR-GTCM), and combined with microscopic tests such as scanning electron microscope (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) and mercury intrusion porosimetry (MIP) for the enhancement mechanism were deeply analysed. The results show that 0.3% BF and 20% GT are the optimal doping amounts for the mechanical response of BFR-GTCM, which especially significantly improves the flexural and crack resistance. In addition, the synergistic effect of basalt fibres and graphite tailings optimizes the spatial structure and pore distribution of the cement matrix and promoted the hydration of the cementitious material, thus improving the mechanical properties of BFR-GTCM. Full article

Recombination is the main driver of bacteriophage evolution. It may serve as a tool for extending the phage host spectrum, which is significant not only for phages’ ecology but also for their utilisation as therapeutic agents of bacterial infections. The aim of this study was to detect the recombination events in the genomes of Litunavirus phages infecting Pseudomonas aeruginosa, and present their impact on phylogenetic relations within this phage group. The phylogenetic analyses involved: the whole-genome, core-genome (Schitoviridae conserved genes), variable genome region, and the whole-genome minus variable region. Interestingly, the recombination events taking place in the putative host recognition region (tail fibre protein gene and the adjacent downstream gene) significantly influenced tree topology, suggesting a strong phylogenetic signal. Our results indicate the recombination between phages from two genera Litunavirus and Luzeptimavirus and demonstrate its influence on phage phylogeny. Full article

ODF1 is a major protein of the accessory fibres of the mammalian sperm tail. In addition, ODF1 is found in the connecting piece, a complex structure located at the posterior end of the nucleus that connects the sperm head and tail. The tight coupling of the sperm head and tail is critical for the progressive motility of the sperm to reach the oocyte for fertilisation. The depletion of ODF1 by homologous recombination in mice led to male infertility. Although sperm tails were present in the epididymis, no intact spermatozoa were found. Instead, the depletion of ODF1 resulted in sperm decapitation, suggesting that ODF1 is essential for the formation of the coupling apparatus and the tight linkage of the sperm head and tail. However, the development of the linkage complex in the absence of ODF1 has never been investigated. Here, I analysed the fine structure of the developing connecting piece by transmission electron microscopy. I show that the connecting piece develops as in wild-type spermatids. Structural abnormalities were not observed when ODF1 was absent. Thus, ODF1 is dispensable for the development of the connecting piece. However, the decapitation of ODF1-deficient spermatozoa indicates that the heads and tails of the spermatozoa are not linked, so that they separate when force is applied. Full article

Wet spinning is an established fibre manufacturing route to realise collagen fibres with preserved triple helix architecture and cell acceptability for applications in biomedical membranes. However, resulting fibres still need to be chemically modified post-spinning to ensure material integrity in physiological media, with inherent risks of alteration of fibre morphology and with limited opportunities to induce fibrillogenesis following collagen fixation in the crosslinked state. To overcome this challenge, we hypothesised that a photoactive type I collagen precursor bearing either single or multiple monomers could be employed to accomplish hierarchically assembled fibres with improved processability, macroscopic properties and nanoscale organisation via sequential wet spinning and UV-curing. In-house-extracted type I rat tail collagen functionalised with both 4-vinylbenzyl chloride (4VBC) and methacrylate residues generated a full hydrogel network following solubilisation in a photoactive aqueous solution and UV exposure, whereby ~85 wt.% of material was retained following 75-day hydrolytic incubation. Wide-angle X-ray diffraction confirmed the presence of typical collagen patterns, whilst an averaged compression modulus and swelling ratio of more than 290 kPa and 1500 wt.% was recorded in the UV-cured hydrogel networks. Photoactive type I collagen precursors were subsequently wet spun into fibres, displaying the typical dichroic features of collagen and regular fibre morphology. Varying tensile modulus (E = 5 ± 1 − 11 ± 4 MPa) and swelling ratio (SR = 1880 ± 200 − 3350 ± 500 wt.%) were measured following post-spinning UV curing and equilibration with phosphate-buffered saline (PBS). Most importantly, 72-h incubation of the wet spun fibres in PBS successfully induced renaturation of collagen-like fibrils, which were fixed following UV-induced network formation. The whole process proved to be well tolerated by cells, as indicated by a spread-like cell morphology following a 48-h culture of L929 mouse fibroblasts on the extracts of UV-cured fibres. Full article

The recognition and binding of host bacteria by bacteriophages is most often enabled by a highly specific receptor–ligand type of interaction, with the receptor-binding proteins (RBPs) of phages being the primary determinants of host specificity. Specifically modifying the RBPs could alter or extend the host range of phages otherwise exhibiting desired phenotypic properties. This study employed two different strategies to reprogram T7 phages ordinarily infecting commensal K12 Escherichia coli strains to infect pathogen-associated K1-capsule-expressing strains. The strategies were based on either plasmid-based homologous recombination or bacteriophage recombineering using electroporated DNA (BRED). Our work pursued the construction of two genetic designs: one replacing the gp17 gene of T7, the other replacing gp11, gp12, and gp17 of T7 with their K1F counterparts. Both strategies displayed successful integration of the K1F sequences into the T7 genome, detected by PCR screening. Multiple methods were utilised to select or enrich for chimeric phages incorporating the K1F gp17 alone, including trxA, host-specificity, and CRISPR-Cas-based selection. Irrespective of the selection method, the above strategy yielded poorly reproducible phage propagation on the new host, indicating that the chimeric phage was less fit than the wild type and could not promote continual autonomous reproduction. Chimeric phages obtained from BRED incorporating gp11-12 and gp17, however, all displayed infection in a 2-stage pattern, indicating the presence of both K1F and T7 phenotypes. This study shows that BRED can be used as a tool to quickly access the potential of new RBP constructs without the need to engineer sustainably replicating phages. Additionally, we show that solely repurposing the primary RBP is, in some cases, insufficient to produce a viable chimeric phage. Full article

With the recent rise in interest in using lytic bacteriophages as therapeutic agents, there is an urgent requirement to understand their fundamental biology to enable the engineering of their genomes. Current methods of phage engineering rely on homologous recombination, followed by a system of selection to identify recombinant phages. For bacteriophage T7, the host genes cmk or trxA have been used as a selection mechanism along with both type I and II CRISPR systems to select against wild-type phage and enrich for the desired mutant. Here, we systematically compare all three systems; we show that the use of marker-based selection is the most efficient method and we use this to generate multiple T7 tail fibre mutants. Furthermore, we found the type II CRISPR-Cas system is easier to use and generally more efficient than a type I system in the engineering of phage T7. These results provide a foundation for the future, more efficient engineering of bacteriophage T7. Full article

Bacteriophages of the significant veterinary pathogen Staphylococcus pseudintermedius are rarely described morphologically and genomically in detail, and mostly include phages of the Siphoviridae family. There is currently no taxonomical classification for phages of this bacterial species. Here we describe a new phage designated vB_SpsS_QT1, which is related to phage 2638A originally described as a Staphylococcus aureus phage. Propagating strain S. aureus 2854 of the latter was reclassified by rpoB gene sequencing as S. pseudintermedius 2854 in this work. Both phages have a narrow but different host range determined on 54 strains. Morphologically, both of them belong to the family Siphoviridae, share the B1 morphotype, and differ from other staphylococcal phage genera by a single long fibre at the terminus of the tail. The complete genome of phage vB_SpsS_QT1 was sequenced with the IonTorrent platform and expertly annotated. Its linear genome with cohesive ends is 43,029 bp long and encodes 60 predicted genes with the typical modular structure of staphylococcal siphophages. A global alignment found the genomes of vB_SpsS_QT1 and 2638A to share 84% nucleotide identity, but they have no significant similarity of nucleotide sequences with other phage genomes available in public databases. Based on the morphological, phylogenetic, and genomic analyses, a novel genus Fibralongavirus in the family Siphoviridae is described with phage species vB_SpsS_QT1 and 2638A. Full article

Two experiments were conducted using Dorper × thin-tailed Han crossbred ewes. In experiment 1, eighteen ewes were randomly assigned to two dietary treatments (a basal diet, or the same basal diet supplemented with 2.0 g tea saponin (TS)/head/day) to investigate the effects of TS supplementation on nutrient digestibility and methane emissions. In experiment 2, six ewes with ruminal cannulae were assigned to the same two dietary treatments as in experiment 1 to investigate the effects of TS supplementation on rumen fermentation and microbial flora. TS supplementation increased the apparent digestibility of organic matter (OM) (p = 0.001), nitrogen (N) (p = 0.036), neutral detergent fibre (NDF) (p = 0.001), and acid detergent fibre (ADF) (p < 0.001). Urinary N (p = 0.001) and fecal N (p = 0.036) output were reduced, and N retention (p = 0.001) and nitrogen retention/nitrogen intake (p = 0.001) were increased. Supplementary TS did not decrease absolute methane emissions (p = 0.519) but decreased methane emissions scaled to metabolic bodyweight by 8.80% (p = 0.006). Ammonia levels decreased (p < 0.001) and total volatile fatty acid levels increased (p = 0.018) in response to TS supplementation. The molar proportion of propionate increased (p = 0.007), whereas the acetate:propionate ratio decreased (p = 0.035). Supplementation with TS increased the population of Fibrobacter succinogenes (p = 0.019), but the population of protozoans tended to decrease (p = 0.054). Supplementation with TS effectively enhanced the apparent digestibility of OM, N, NDF, and ADF, and decreased methane emissions scaled to metabolic bodyweight. Full article

Spider aciniform (wrapping) silk is a remarkable fibrillar biomaterial with outstanding mechanical properties. It is a modular protein consisting, in Argiope trifasciata, of a core repetitive domain of 200 amino acid units (W units). In solution, the W units comprise a globular folded core, with five α-helices, and disordered tails that are linked to form a ~63-residue intrinsically disordered linker in concatemers. Herein, we present nuclear magnetic resonance (NMR) spectroscopy-based ¹⁵N spin relaxation analysis, allowing characterization of backbone dynamics as a function of residue on the ps–ns timescale in the context of the single W unit (W₁) and the two unit concatemer (W₂). Unambiguous mapping of backbone dynamics throughout W₂ was made possible by segmental NMR active isotope-enrichment through split intein-mediated trans-splicing. Spectral density mapping for W₁ and W₂ reveals a striking disparity in dynamics between the folded core and the disordered linker and tail regions. These data are also consistent with rotational diffusion behaviour where each globular domain tumbles almost independently of its neighbour. At a localized level, helix 5 exhibits elevated high frequency dynamics relative to the proximal helix 4, supporting a model of fibrillogenesis where this helix unfolds as part of the transition to a mixed α-helix/β-sheet fibre. Full article

Bombus terrestris, commonly known as the buff-tailed bumblebee, is native to Europe, parts of Africa and Asia. It is commercially bred for use as a pollinator of greenhouse crops. Larvae pupate within a silken cocoon that they construct from proteins produced in modified salivary glands. The amino acid composition and protein structure of hand drawn B. terrestris, silk fibres was investigated through the use of micro-Raman spectroscopy. Spectra were obtained from single fibres drawn from the larvae salivary gland at a rate of 0.14 cm/s. Raman spectroscopy enabled the identification of poly(alanine), poly(alanine-glycine), phenylalanine, tryptophan, and methionine, which is consistent with the results of amino acid analysis. The dominant protein conformation was found to be coiled coil (73%) while the β-sheet content of 10% is, as expected, lower than those reported for hornets and ants. Polarized Raman spectra revealed that the coiled coils were highly aligned along the fibre axis while the β-sheet and random coil components had their peptide carbonyl groups roughly perpendicular to the fibre axis. The protein orientation distribution is compared to those of other natural and recombinant silks. A structural model for the B. terrestris silk fibre is proposed based on these results. Full article

Bacteriophage T5, a Siphovirus belonging to the order Caudovirales, has a flexible, three-fold symmetric tail, to which three L-shaped fibres are attached. These fibres recognize oligo-mannose units on the bacterial cell surface prior to infection and are composed of homotrimers of the pb1 protein. Pb1 has 1396 amino acids, of which the carboxy-terminal 133 residues form a trimeric intra-molecular chaperone that is auto-proteolyzed after correct folding. The structure of a trimer of residues 970–1263 was determined by single anomalous dispersion phasing using incorporated selenomethionine residues and refined at 2.3 Å resolution using crystals grown from native, methionine-containing, protein. The protein inhibits phage infection by competition. The phage-distal receptor-binding domain resembles a bullet, with the walls formed by partially intertwined beta-sheets, conferring stability to the structure. The fold of the domain is novel and the topology unique to the pb1 structure. A site-directed mutant (Ser1264 to Ala), in which auto-proteolysis is impeded, was also produced, crystallized and its 2.5 Å structure solved by molecular replacement. The additional chaperone domain (residues 1263–1396) consists of a central trimeric alpha-helical coiled-coil flanked by a mixed alpha-beta domain. Three long beta-hairpin tentacles, one from each chaperone monomer, extend into long curved grooves of the bullet-shaped domain. The chaperone-containing mutant did not inhibit infection by competition. Full article