The Complexity of the Ovine and Caprine Keratin-Associated Protein Genes

Sheep (Ovis aries) and goats (Capra hircus) have, for more than a millennia, been a source of fibres for human use, be it for use in clothing and furnishings, for insulation, for decorative and ceremonial purposes, or for combinations thereof. While use of these natural fibres has in some respects been superseded by the use of synthetic and plant-based fibres, increased accounting for the carbon and water footprint of these fibres is creating a re-emergence of interest in fibres derived from sheep and goats. The keratin-associated proteins (KAPs) are structural components of wool and hair fibres, where they form a matrix that cross-links with the keratin intermediate filaments (KIFs), the other main structural component of the fibres. Since the first report of a complete KAP protein sequence in the late 1960s, considerable effort has been made to identify the KAP proteins and their genes in mammals, and to ascertain how these genes and proteins control fibre growth and characteristics. This effort is ongoing, with more and more being understood about the structure and function of the genes. This review consolidates that knowledge and suggests future directions for research to further our understanding.

Comprehensive multidimensional study of the self-assembly properties of a three residue substituted β3 oligoamide

Substituted β3 oligoamides form a unique self-assembling system where each monomer folds into a helix containing approximately three β3 amino acids per turn, yielding a geometrically well-defined cylindrical building block that, when N-acylated, is able to self-assemble head-to-tail into nanorods that can reach several 100 μm length. It was shown in previous works that self-assembly can be achieved with a three residue long oligoamide as well that lacks any intramolecular H-bonds, yet it crystallizes in a helix-like conformation. The self-assembly properties of these small oligoamides are however elusive, suggesting a more complex system than the self-assembly of the H-bond stabilized helical monomers. Here we focus on the self-assembly behaviour of a three residue oligoamide, Ac-β3[LIA] where the letters denote the side chain of the analogous α amino acid. Ac-β3[LIA] can yield highly inhomogeneous suspensions in water with a broad range of large fibrous structures that seem to be very stable, yet occasionally fibre growth is only observed upon heating. The small size of the monomer suggests a highly dynamic equilibrium yet all previous attempts failed to clearly identify low molecular weight species. Therefore a special methodology was employed in this study to characterize the suspensions at different size ranges: SANS that is optimal to measure the small oligomers and cross sectional diameter of the assemblies, DLS that is sensitive to the large populations and therefore the length of the superstructures, and NMR that is sensitive to monomeric and small oligomeric form, in conjunction with IR spectroscopy to probe the folding and AFM to image the morphology of the assemblies. Temperature ramping was used to perturb the system to probe the dynamicity of the self-assembly. It was found that the anomalous self-assembly behaviour of Ac-β3[LIA] is caused by its two stable conformations, a helix-building “horseshoe” fold and a linear conformer. The latter is exclusively found in monomeric form in solution whereas the horseshoe fold is stable in solid phase and in fibrous assemblies. Small oligomers were absent. Thus the self-assembly of Ac-β3[LIA] is arrested by the activation energy need of the conformation change; fibre growth might be triggered by conditions that allow increased conformational freedom of the monomers. This observation may be used to develop strategies for controlled switchable self-assembly.

Fibrinogen αC-subregions critically contribute blood clot fibre growth, mechanical stability and resistance to fibrinolysis

Fibrinogen is essential for blood coagulation. The C-terminus of the fibrinogen α-chain (αC-region) is composed of an αC-domain and αC-connector. Two recombinant fibrinogen variants (α390 and α220) were produced to investigate the role of subregions in modulating clot stability and resistance to lysis. The α390 variant, truncated before the αC-domain, produced clots with a denser structure and thinner fibres. In contrast, the α220 variant, truncated at the start of the αC-connector, produced clots that were porous with short, stunted fibres and visible fibre ends. These clots were mechanically weak and susceptible to lysis. Our data demonstrate differential effects for the αC-subregions in fibrin polymerisation, clot mechanical strength, and fibrinolytic susceptibility. Furthermore, we demonstrate that the αC-subregions are key for promoting longitudinal fibre growth. Together, these findings highlight critical functions of the αC-subregions in relation to clot structure and stability, with future implications for development of novel therapeutics for thrombosis.

Fibrinogen αC-subregions critically contribute blood clot fibre growth, mechanical stability and resistance to fibrinolysis

Fibrinogen is essential for blood coagulation. The C-terminus of the fibrinogen α-chain (αC-region) is composed of an αC-domain and αC-connector. Two recombinant fibrinogen variants (α390 and α220) were produced to investigate the role of subregions in modulating clot stability and resistance to lysis. The α390 variant, truncated before the αC-domain, produced clots with a denser structure and thinner fibres. In contrast, the α220 variant, truncated at the start of the αC-connector, produced clots that were porous with short stunted fibres and visible fibre ends. These clots were mechanically weak and susceptible to lysis. Our data demonstrate differential effects for the αC-subregions in fibrin polymerisation, clot mechanical strength, and fibrinolytic susceptibility. Furthermore, we demonstrate that the αC-subregions are key for promoting longitudinal fibre growth. Together, these findings highlight critical functions of the αC-subregions in relation to clot structure and stability, with future implications for development of novel therapeutics for thrombosis.

Effects of dietary methionine on growth performance, muscle nutritive deposition, muscle fibre growth and type I collagen synthesis of on-growing grass carp (Ctenopharyngodon idella)

In the current research, a 60-day experiment was conducted with the purpose of exploring the impacts of methionine on growth performance, muscle nutritive deposition, muscle fibre growth and type I collagen synthesis as well as related signaling pathway. Six diets (iso-nitrogenous) differing in methionine concentrations (2.54, 4.85, 7.43, 10.12, 12.40 and 15.11 g kg-1 diets) were fed to 540 grass carp (178.47 ± 0.36 g). Results showed (P < 0.05) that, compared with methionine deficiency, optimal level of dietary methionine (1) increased feed intake (FI), feed efficiency (FE), specific growth rate (SGR) and percentage weight gain (PWG); (2) increased fish muscle protein, lipid and free amino acid contents, and improved fish muscle fatty acid profile as well as increased protein content in part associated with TORC1/S6K1 signaling pathway; (3) increased the frequency distribution of muscle fibre with >50 µm of diameter; (4) increased type I collagen synthesis partly related to TGF-β1/Smads and CK2/TORC1 signaling pathways. In conclusion, dietary methionine improved muscle growth, which might be due to the regulation of muscle nutritive deposition, and muscle fibre growth and type I collagen synthesis related signal molecules. Finally, according to PWG and muscle collagen content, the methionine requirements for on-growing grass carp (178-626 g) were estimated to be 9.56 g kg-1 diet (33.26 g kg-1 protein of diet) and 9.28 g kg-1 diet (32.29 g kg-1 of dietary protein), respectively.

Nogo-A Beyond the RhoA/ROCK Pathway – Novel Components of Intracellular Nogo-A Signaling Cascades

Nogo-A is a well-characterized myelin-associated membrane protein that restricts fibre growth and the regenerative capacity of the adult central nervous system after injury. To date Nogo-A post-receptor signalling pathway research focused on the RhoA/ROCK cascade, which can lead to growth cone collapse and neurite retraction. Much less is known about continued intracellular Nogo-A signalling mediating long-term neurite outgrowth inhibition resulting from transcriptional and translational changes. Here, we propose a simple but highly reproducible in vitro assay to study Nogo-A related signaling and neurite outgrowth inhibition in general. Furthermore, we identified ERK1/2 as downstream effector of Nogo-A, partially mediating its neurite outgrowth inhibition. We describe ERK1/2 dependent changes of translational events such as elevation of RhoA levels within the growth cone, which may potentiate the cells’ responses to Nogo-A. We also observed Nogo-A dependent upregulation of the JAK/STAT pathway inhibitors SOCS3 and KLF4 and downregulation of insulin mediated phosphorylation of AKT, indicating direct negative crosstalk between Nogo-A signalling and the growth promoting JAK/STAT and AKT/mTORC1 pathways.

Eriodictyol can modulate cellular auxin gradients to efficiently promote in vitro cotton fibre development

Background Flavonoids have essential roles in flower pigmentation, fibre development and disease resistance in cotton. Previous studies show that accumulation of naringenin in developing cotton fibres significantly affects fibre growth. This study focused on determining the effects of the flavonoids naringenin, dihydrokaempferol, dihydroquerectin and eriodictyol on fibre development in an in vitro system. Results 20 μM eriodictyol treatment produced a maximum fibre growth, in terms of fibre length and total fibre units. To gain insight into the associated transcriptional regulatory networks, RNA-seq analysis was performed on eriodictyol-treated elongated fibres, and computational analysis of differentially expressed genes revealed that carbohydrate metabolism and phytohormone signaling pathways were differentially modulated. Eriodictyol treatment also promoted the biosynthesis of quercetin and dihydroquerectin in ovules and elongating fibres through enhanced expression of genes encoding chalcone isomerase, chalcone synthase and flavanone 3-hydroxylase. In addition, auxin biosynthesis and signaling pathway genes were differentially expressed in eriodictyol-driven in vitro fibre elongation. In absence of auxin, eriodictyol predominantly enhanced fibre growth when the localized auxin gradient was disrupted by the auxin transport inhibitor, triiodobenzoic acid. Conclusion Eriodictyol was found to significantly enhance fibre development through accumulating and maintaining the temporal auxin gradient in developing unicellular cotton fibres.

Origin of spherulitic and cone-in-cone concretions in Cambro-Ordovician black shales, St Lawrence Estuary, Quebec, Canada

Spherulitic concretions are very rare among carbonate concretions that generally consist of micritic carbonate. The occurrence of spherulitic concretions in Cambro-Ordovician black shales of unknown stratigraphic age on a mid-channel island in the St Lawrence Estuary in Quebec is a new example in addition to only three hitherto reported occurrences of spherulitic carbonate concretions. Their origin is still poorly understood. These concretions occur in close association with, and show various transitions to, cone-in-cone structure. The spherules, measuring 0.5 to 12 mm in diameter, consist of intergrown fine fibres of ferroan calcite and quartzine, pointing to the formation of the concretions below the sulfate-reduction zone. A phenomenological theory of spherulitic crystallization relates the thickness δ of an impurity-rich layer in front of impurity-rejecting growing crystals to the impurity-diffusion coefficient D and the growth velocity G of the crystal by δ = D/G. In spherulite-forming environments, extremely small values of δ (in the order of <10−4 cm) in conjunction with cellulation lead to spherulitic fibre growth. The theory of spherulitic crystallization is here applied to sedimentary deposits for the first time. The intimate association of calcite and quartzine in the concretions requires a chemical change from alkaline to acidic conditions, which occurs below the carbonate-reduction zone owing to the dissolution of sponge spicules or radiolarians. The transition from spherulite to the silica-free cone-in-cone structure occurs when the silica reservoir that acted as an impurity is exhausted in the crystallization process.