Rotary Jet Spinning of Polymer Fibres

Polymeric nanofibres can be produced from a variety of methods such as electrospinning and melt blowing, with fibres being produced having applications in many sectors such as biomedicine, composites and filtration. Existing methods are not however capable of producing nanofibres to commercial volumes in an energy efficient way. In this research we investigate a new method of producing nanofibres, namely Rotary Jet Spinning (RJS), which is a relatively new method of fibre production similar to candyfloss production, where centrifugal forces are used to expel jets of polymer from a state of melt or solution in order to produce polymeric fibres. We investigate this method in detail, initially concentrating on the comparison between electrospinning and RJS. Firstly, it was found that electrospinning produced slightly smaller fibre diameters compared to RJS over a broader range of solution concentrations. Secondly, the ability to produce high modulus fibres was investigated by means of an imidization technique, where polyamic acid solution was produced and spun into fibres before conversion to a co-polyimide fibre with an elastic modulus of around 40 GPa. In the third experimental chapter, the viscosity reliability of the RJS process was evaluated by means of computational fluid dynamics simulations, where it was shown that low viscosity (1-10 Pa.s) Newtonian fluids are required to establish fibre production. For fluids with lower viscosities, beading occurred in solution spinning and droplets were produced from melt spinning. Viscosities higher than the recommended value resulted in blockage, with no fibres being produced from either method. Lastly, the production of ceramic fibres was evaluated to establish the ability of the RJS process to produce a ceramic nanofibre. Fibres on the nanoscale were not achieved, however a variation in solvent volatility and crosslinking time were factors in fibre diameter reduction, with solvent variations highlighting the potential of this process to achieve the required fibre size from RJS and thereby demonstrating this technology as a viable option for high volume fibre production.EPSRC grant number 150219

PA6 nanofibre production: A comparison between rotary jet spinning and electrospinning

© 2018 by the authors. Polymer nanofibres are created from many different techniques, with varying rates of production. Rotary jet spinning is a relatively new technique for making nanofibres from both polymer solutions and melt. With electrospinning being by far the most widespread processing method for polymer nanofibres, we performed a direct comparison of polyamide 6 (PA6) nanofibre production between these two methods. It was found that electrospinning produced slightly smaller-diameter fibres, which scaled with a decrease in solution viscosity. In comparison, rotary jet spun fibres could be produced from a reduced range of polymer concentrations and exhibited therefore slightly larger diameters with greater variation. Crystallinity of the fibres was also compared between the two techniques and the bulk polymer, which showed a decrease in crystallinity compared to bulk PA6

Calpains Mediate Integrin Attachment Complex Maintenance of Adult Muscle in Caenorhabditis elegans

Two components of integrin containing attachment complexes, UNC-97/PINCH and UNC-112/MIG-2/Kindlin-2, were recently identified as negative regulators of muscle protein degradation and as having decreased mRNA levels in response to spaceflight. Integrin complexes transmit force between the inside and outside of muscle cells and signal changes in muscle size in response to force and, perhaps, disuse. We therefore investigated the effects of acute decreases in expression of the genes encoding these multi-protein complexes. We find that in fully developed adult Caenorhabditis elegans muscle, RNAi against genes encoding core, and peripheral, members of these complexes induces protein degradation, myofibrillar and mitochondrial dystrophies, and a movement defect. Genetic disruption of Z-line– or M-line–specific complex members is sufficient to induce these defects. We confirmed that defects occur in temperature-sensitive mutants for two of the genes: unc-52, which encodes the extra-cellular ligand Perlecan, and unc-112, which encodes the intracellular component Kindlin-2. These results demonstrate that integrin containing attachment complexes, as a whole, are required for proper maintenance of adult muscle. These defects, and collapse of arrayed attachment complexes into ball like structures, are blocked when DIM-1 levels are reduced. Degradation is also blocked by RNAi or drugs targeting calpains, implying that disruption of integrin containing complexes results in calpain activation. In wild-type animals, either during development or in adults, RNAi against calpain genes results in integrin muscle attachment disruptions and consequent sub-cellular defects. These results demonstrate that calpains are required for proper assembly and maintenance of integrin attachment complexes. Taken together our data provide in vivo evidence that a calpain-based molecular repair mechanism exists for dealing with attachment complex disruption in adult muscle. Since C. elegans lacks satellite cells, this mechanism is intrinsic to the muscles and raises the question if such a mechanism also exists in higher metazoans

Diapause as escape strategy to exposure to toxicants: response of Brachionus calyciforus to arsenic

Invertebrate organisms commonly respond to environmental fluctuation by entering diapause. Production of diapause in monogonont rotifers involves a previous switch from asexual to partial sexual reproduction. Although zooplankton have been used in ecotoxicological assays, often their true vulnerability to toxicants is underestimated by not incorporating the sexual phase. We experimentally analyzed traits involved in sexual reproduction and diapause in the cyclically parthenogenetic freshwater rotifer, Brachionus calyciflorus, exposed to arsenic, a metalloid naturally found in high concentrations in desert zones, focusing on the effectiveness of diapause as an escape response in the face of an adverse condition. Addition of sublethal concentrations of arsenic modified the pattern of diapause observed in the rotifer: investment in diapause with arsenic addition peaked earlier and higher than in non-toxicant conditions, which suggests that sexual investment could be enhanced in highly stressed environmental conditions by increased responsiveness to stimulation. Nevertheless, eggs produced in large amount with arsenic, were mostly low quality, and healthy-looking eggs had lower hatching success, therefore it is unclear whether this pattern is optimum in an environment with arsenic, or if rather arsenic presence in water bodies disturbs the optimal allocation of offspring entering diapause. We observed high accumulation of arsenic in organisms exposed to constant concentration after several generations, which suggests that arsenic may be accumulated transgenerationally. The sexual phase in rotifers may be more sensitive to environmental conditions than the asexual one, therefore diapause attributes should be considered in ecotoxicological assessment because of its ecological and evolutionary implications on lakes biodiversity

Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells

The original publication is available at http:/www.plosone.orgBackground: This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first. Methodology/Principal Findings: In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions. Conclusions: The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening. © 2011 Sharathchandra et al.Publishers' Versio

Phylogeny of Parasitic Parabasalia and Free-Living Relatives Inferred from Conventional Markers vs. Rpb1, a Single-Copy Gene

Parabasalia are single-celled eukaryotes (protists) that are mainly comprised of endosymbionts of termites and wood roaches, intestinal commensals, human or veterinary parasites, and free-living species. Phylogenetic comparisons of parabasalids are typically based upon morphological characters and 18S ribosomal RNA gene sequence data (rDNA), while biochemical or molecular studies of parabasalids are limited to a few axenically cultivable parasites. These previous analyses and other studies based on PCR amplification of duplicated protein-coding genes are unable to fully resolve the evolutionary relationships of parabasalids. As a result, genetic studies of Parabasalia lag behind other organisms.Comparing parabasalid EF1α, α-tubulin, enolase and MDH protein-coding genes with information from the Trichomonas vaginalis genome reveals difficulty in resolving the history of species or isolates apart from duplicated genes. A conserved single-copy gene encodes the largest subunit of RNA polymerase II (Rpb1) in T. vaginalis and other eukaryotes. Here we directly sequenced Rpb1 degenerate PCR products from 10 parabasalid genera, including several T. vaginalis isolates and avian isolates, and compared these data by phylogenetic analyses. Rpb1 genes from parabasalids, diplomonads, Parabodo, Diplonema and Percolomonas were all intronless, unlike intron-rich homologs in Naegleria, Jakoba and Malawimonas.The phylogeny of Rpb1 from parasitic and free-living parabasalids, and conserved Rpb1 insertions, support Trichomonadea, Tritrichomonadea, and Hypotrichomonadea as monophyletic groups. These results are consistent with prior analyses of rDNA and GAPDH sequences and ultrastructural data. The Rpb1 phylogenetic tree also resolves species- and isolate-level relationships. These findings, together with the relative ease of Rpb1 isolation, make it an attractive tool for evaluating more extensive relationships within Parabasalia

Fermitins, the orthologs of mammalian Kindlins, regulate the development of a functional cardiac syncytium in Drosophila melanogaster.

The vertebrate Kindlins are an evolutionarily conserved family of proteins critical for integrin signalling and cell adhesion. Kindlin-2 (KIND2) is associated with intercalated discs in mice, suggesting a role in cardiac syncytium development; however, deficiency of Kind2 leads to embryonic lethality. Morpholino knock-down of Kind2 in zebrafish has a pleiotropic effect on development that includes the heart. It therefore remains unclear whether cardiomyocyte Kind2 expression is required for cardiomyocyte junction formation and the development of normal cardiac function. To address this question, the expression of Fermitin 1 and Fermitin 2 (Fit1, Fit2), the two Drosophila orthologs of Kind2, was silenced in Drosophila cardiomyocytes. Heart development was assessed in adult flies by immunological methods and videomicroscopy. Silencing both Fit1 and Fit2 led to a severe cardiomyopathy characterised by the failure of cardiomyocytes to develop as a functional syncytium and loss of synchrony between cardiomyocytes. A null allele of Fit1 was generated but this had no impact on the heart. Similarly, the silencing of Fit2 failed to affect heart function. In contrast, the silencing of Fit2 in the cardiomyocytes of Fit1 null flies disrupted syncytium development, leading to severe cardiomyopathy. The data definitively demonstrate a role for Fermitins in the development of a functional cardiac syncytium in Drosophila. The findings also show that the Fermitins can functionally compensate for each other in order to control syncytium development. These findings support the concept that abnormalities in cardiomyocyte KIND2 expression or function may contribute to cardiomyopathies in humans

Skew Calabi-Yau algebras and homological identities

A skew Calabi-Yau algebra is a generalization of a Calabi-Yau algebra which allows for a non-trivial Nakayama automorphism. We prove three homological identities about the Nakayama automorphism and give several applications. The identities we prove show (i) how the Nakayama automorphism of a smash product algebra A#. H is related to the Nakayama automorphisms of a graded skew Calabi-Yau algebra A and a finite-dimensional Hopf algebra H that acts on it; (ii) how the Nakayama automorphism of a graded twist of A is related to the Nakayama automorphism of A; and (iii) that the Nakayama automorphism of a skew Calabi-Yau algebra A has trivial homological determinant in case A is noetherian, connected graded, and Koszul. © 2014 Elsevier Inc

Skew Calabi-Yau triangulated categories and frobenius ext-algebras

We investigate conditions that are sufficient to make the Extalgebra of an object in a (triangulated) category into a Frobenius algebra, and compute the corresponding Nakayama automorphism. As an application, we prove the conjecture that hdet(μA) = 1 for any noetherian Artin-Schelter regular (hence skew Calabi-Yau) algebra A