Nanostructures of etherified arabinoxylans and the effect of arabinose content on material properties.

To further our understanding of a thermoplastic arabinoxylan (AX) material obtained through an oxidation-reduction-etherification pathway, the role of the initial arabinose:xylose ratio on the material properties was investigated. Compression molded films with one molar substitution of butyl glycidyl ether (BGE) showed markedly different tensile behaviors. Films made from low arabinose AX were less ductile, while those made from high arabinose AX exhibited elastomer-like behaviors. X-ray scattering confirmed the presence of nanostructure formation resulting in nano-domains rich in either AX or BGE, from side chain grafting. The scattering data showed variations in the presence of ordered structures, nano-domain sizes and their temperature response between AX with different arabinose contents. In dynamic mechanical testing, three transitions were observed at approximately -90 °C, -50 °C and 80 °C, with a correlation between samples with more structured nano-domains and those with higher onset transition temperatures and lower storage modulus decrease. The mechanical properties of the final thermoplastic AX material can therefore be tuned by controlling the composition of the starting material.

Alpha-secretase dependent nuclear localization of the amyloid-ß precursor protein-binding protein Fe65 promotes DNA repair.

Fe65 is a brain enriched adaptor protein involved in various cellular processes, including actin cytoskeleton regulation, DNA repair and transcription. A well-studied interacting partner of Fe65 is the transmembrane amyloid-ß precursor protein (APP), which can undergo regulated intramembrane proteolysis (RIP). Following ß- and γ-secretase-mediated RIP, the released APP intracellular domain (AICD) together with Fe65 can translocate to the nucleus and regulate transcription. In this study, we investigated if Fe65 nuclear localization can also be regulated by different α-secretases, also known to participate in RIP of APP and other transmembrane proteins. We found that in both Phorbol 12-myristate 13-acetate and all-trans retinoic acid differentiated neuroblastoma cells a strong negative impact on Fe65 nuclear localization, equal to the effect observed upon γ-secretase inhibition, could be detected following inhibition of all three (ADAM9, ADAM10 and ADAM17) α-secretases. Moreover, using the comet assay and analysis of Fe65 dependent DNA repair associated posttranslational modifications of histones, we could show that inhibition of α-secretase-mediated Fe65 nuclear translocation resulted in impaired capacity of the cells to repair DNA damage. Taken together this suggests that α-secretase processing of APP and/or other Fe65 interacting transmembrane proteins play an important role in regulating Fe65 nuclear translocation and DNA repair.

Ethylcellulose-stabilized fat-tissue phantom for quality assurance in clinical hyperthermia.

BACKGROUND: Phantoms accurately mimicking the electromagnetic and thermal properties of human tissues are essential for the development, characterization, and quality assurance (QA) of clinically used equipment for Hyperthermia Treatment (HT). Currently, a viable recipe for a fat equivalent phantom is not available, mainly due to challenges in the fabrication process and fast deterioration. MATERIALS AND METHODS: We propose to employ a glycerol-in-oil emulsion stabilized with ethylcellulose to develop a fat-mimicking material. The dielectric, rheological, and thermal properties of the phantom have been assessed by state-of-the-art measurement techniques. The full-size phantom was then verified in compliance with QA guidelines for superficial HT, both numerically and experimentally, considering the properties variability. RESULTS: Dielectric and thermal properties were proven equivalent to fat tissue, with an acceptable variability, in the 8 MHz to 1 GHz range. The rheology measurements highlighted enhanced mechanical stability over a large temperature range. Both numerical and experimental evaluations proved the suitability of the phantom for QA procedures. The impact of the dielectric property variations on the temperature distribution has been numerically proven to be limited (around 5%), even if higher for capacitive devices (up to 20%). CONCLUSIONS: The proposed fat-mimicking phantom is a good candidate for hyperthermia technology assessment processes, adequately representing both dielectric and thermal properties of the human fat tissue while maintaining structural stability even at elevated temperatures. However, further experimental investigations on capacitive heating devices are necessary to better assess the impact of the low electrical conductivity values on the thermal distribution.

Morphology and swelling of thin films of dialcohol xylan.

Polysaccharides are excellent network formers and are often processed into films from water solutions. Despite being hydrophilic polysaccharides, the typical xylans liberated from wood are sparsely soluble in water. We have previously suggested that an additional piece to the solubilization puzzle is modification of the xylan backbone via oxidative cleavage of the saccharide ring. Here, we demonstrate the influence of the degree of modification, i.e., degree of oxidation (DO) on xylan solubilization and consequent film formation and stability. Oxidized and reduced wood xylans (i.e., dialcohol xylans) with the highest DO (77 %) within the series exhibited the smallest hydrodynamic diameter (dh) of 60 nm in dimethylsulfoxide (DMSO). We transferred the modified xylans into films credit to their established solubility and then quantified the film water interactions. Dialcohol xylans with intermediate DOs (42 and 63 %) did not form continuous films. The films swelled slightly when subjected to humidity. However, the film with the highest DO demonstrated a significant moisture uptake that depended on the film mass and was not observed with the other modified grades or with unmodified xylan.

Potential of Wood Hemicelluloses and Their Derivates as Food Ingredients.

A holistic utilization of all lignocellulosic wood biomass, instead of the current approach of using only the cellulose fraction, is crucial for the efficient, ecological, and economical use of the forest resources. Use of wood constituents in the food and feed sector is a potential way of promoting the global economy. However, industrially established food products utilizing such components are still scarce, with the exception of cellulose derivatives. Hemicelluloses that include xylans and mannans are major constituents of wood. The wood hemicelluloses are structurally similar to hemicelluloses from crops, which are included in our diet, for example, as a part of dietary fibers. Hence, structurally similar wood hemicelluloses have the potential for similar uses. We review the current status and future potential of wood hemicelluloses as food ingredients. We include an inventory of the extraction routes of wood hemicelluloses, their physicochemical properties, and some of their gastrointestinal characteristics, and we also consider the regulatory route that research findings need to follow to be approved for food solutions, as well as the current status of the wood hemicellulose applications on that route.

Lingonberry (Vaccinium vitis-idaea) press-cake as a new processing aid during isolation of protein from herring (Clupea harengus) co-products.

High acid-consumption and lipid oxidation are challenges when recovering functional proteins from herring co-products via pH-shift-processing. Here, lingonberry press-cake (LP), which is abundant in organic acids and phenolics, was added to alkali-solubilized herring-co-product-proteins (2.5-30 % LP per dry weight) aiming to aid protein precipitation, save hydrochloric acid (HCl) and provide oxidative stability. The results revealed 5-30 % LP addition reduced HCl-consumption by 13-61 % and 19-79 % when precipitating proteins at pH 5.5 and 6.5, respectively. Higher LP% decreased protein content and lightness of protein isolates but raised the lipid content. Precipitation at pH 6.5 used less acid, reduced total protein yield and raised moisture content and darkness of isolates. Contrary to controls, lipid oxidation-derived volatiles did not develop in protein isolates precipitated with 10 % and 30 % LP, neither during the process itself nor during 21 days on ice. Altogether, LP was identified as a promising all-natural processing-aid to use during herring protein isolation.

Radial discharge high shear homogenization and ultrasonication assisted pH-shift processing of herring co-products with antioxidant-rich materials for maximum protein yield and functionality.

Cross-processing herring co-products with antioxidant-rich helpers including lingonberry-press-cake, shrimp-shells and seaweed was reported to mitigate lipid oxidation but reduce protein yield. Here, four strategies were used to counteract such yield-reduction; optimizing solubilization/precipitation pH, increasing raw-material-to-water-ratio, replacing single-stage-toothed- by radial-discharge- high-shear-mechanical-homogenization (RD-HSMH) and ultrasonication (US). The effects of RD-HSMH and US on lipid oxidation, protein structural and functional properties were studied. Combining four strategies improved total protein yield by 5-12 %, depending on helper type. More than the confirmed antioxidant effects, cross-processing also improved protein water solubility and emulsification activity but reduced gelation properties. RD-HSMH generally improved protein emulsifying and gelation properties but reduced protein water solubility. US reduced protein water solubility and gelation properties. Altogether, it was recommended for all helpers to increase solubilization pH to 12 and raw-material-to-water-ratio to 1:6 followed by RD-HSMH at 8000 rpm for 90 s, aiming for maximum protein yield and emulsifying and gelation properties.

Key Modulators of the Stress Granule Response TIA1, TDP-43, and G3BP1 Are Altered by Polyglutamine-Expanded ATXN7.

Spinocerebellar ataxia type 7 (SCA7) and other polyglutamine (polyQ) diseases are caused by expansions of polyQ repeats in disease-specific proteins. Aggregation of the polyQ proteins resulting in various forms of cellular stress, that could induce the stress granule (SG) response, is believed to be a common pathological mechanism in these disorders. SGs can contribute to cell survival but have also been suggested to exacerbate disease pathology by seeding protein aggregation. In this study, we show that two SG-related proteins, TDP-43 and TIA1, are sequestered into the aggregates formed by polyQ-expanded ATXN7 in SCA7 cells. Interestingly, mutant ATXN7 also localises to induced SGs, and this association altered the shape of the SGs. In spite of this, neither the ability to induce nor to disassemble SGs, in response to arsenite stress induction or relief, was affected in SCA7 cells. Moreover, we could not observe any change in the number of ATXN7 aggregates per cell following SG induction, although a small, non-significant, increase in total aggregated ATXN7 material could be detected using filter trap. However, mutant ATXN7 expression in itself increased the speckling of the SG-nucleating protein G3BP1 and the SG response. Taken together, our results indicate that the SG response is induced, and although some key modulators of SGs show altered behaviour, the dynamics of SGs appear normal in the presence of mutant ATXN7.

Modification of xylan via an oxidation-reduction reaction.

Xylan is a biopolymer readily available from forest resources. Various modification methods, including oxidation with sodium periodate, have been shown to facilitate the engineering applications of xylan. However, modification procedures are often optimized for semicrystalline high molecular weight polysaccharide cellulose rather than for lower molecular weight and amorphous polysaccharide xylan. This paper elucidates the procedure for the periodate oxidation of xylan into dialdehyde xylan and its further reduction into a dialcohol form and is focused on the modification work up. The oxidation-reduction reaction decreased the molecular weight of xylan while increased the dispersity more than 50%. Unlike the unmodified xylan, all the modified grades could be solubilized in water, which we see essential for facilitating the future engineering applications of xylan. The selection of quenching and purification procedures and pH-adjustment of the reduction step had no significant effect on the degree of oxidation, molecular weight and only a minor effect on the hydrodynamic radius in water. Hence, it is possible to choose the simplest oxidation-reduction route without time consuming purification steps within the sequence.

Effect of physicochemical properties, pre-processing, and extraction on the functionality of wheat bran arabinoxylans in breadmaking - A review.

Arabinoxylan (AX) is an abundant hemicellulose in wheat bran and an important functional component in bakery products. This review compares preprocessing and extraction methods, and evaluates their effect on AX properties and functionality as a bread ingredient. The extraction process results in AX isolates or concentrates with varying molecular characteristics, indicating that the process can be adjusted to produce AX with targeted functionality. AX functionality in bread seems to depend on AX properties but also on AX addition level and interactions with other components. This review suggests that the use of AX with tailored properties together with properly optimized baking process could help increasing the amount of added fiber in bread while maintaining or even improving bread quality.

Monitoring of Chromatin Organization at the Nuclear Pore Complex, Inner Nuclear Membrane, and Nuclear Interior in Live Cells by Fluorescence Ratiometric Imaging of Chromatin (FRIC).

The image analysis tool FRIC (Fluorescence Ratiometric Imaging of Chromatin) quantitatively monitors dynamic spatiotemporal distribution of euchromatin and total chromatin in live cells. A vector (pTandemH) assures stoichiometrically constant expression of the histone variants Histone 3.3 and Histone 2B, fused to EGFP and mCherry, respectively. Quantitative ratiometric (H3.3/H2B) imaging displayed a concentrated distribution of heterochromatin in the periphery of U2OS cell nuclei. As a proof of concept, peripheral heterochromatin responded to experimental manipulation of histone acetylation as well as expression of the mutant lamin A protein "progerin," which causes Hutchinson-Gilford Progeria Syndrome. In summary FRIC is versatile, unbiased, robust, requires a minimum of experimental steps and is suitable for screening purposes.

Impact of Glucose on the Nanostructure and Mechanical Properties of Calcium-Alginate Hydrogels.

Alginate is a polysaccharide obtained from brown seaweed that is widely used in food, pharmaceutical, and biotechnological applications due to its versatility as a viscosifier and gelling agent. Here, we investigated the influence of the addition of glucose on the structure and mechanical properties of alginate solutions and calcium-alginate hydrogels produced by internal gelation through crosslinking with Ca2+. Using 1H low-field nuclear magnetic resonance (NMR) and small angle neutron scattering (SANS), we showed that alginate solutions at 1 wt % present structural heterogeneities at local scale whose size increases with glucose concentration (15-45 wt %). Remarkably, the molecular conformation of alginate in the gels obtained from internal gelation by Ca2+ crosslinking is similar to that found in solution. The mechanical properties of the gels evidence an increase in gel strength and elasticity upon the addition of glucose. The fitting of mechanical properties to a poroelastic model shows that structural changes within solutions prior to gelation and the increase in solvent viscosity contribute to the gel strength. The nanostructure of the gels (at local scale, i.e., up to few hundreds of Å) remains unaltered by the presence of glucose up to 30 wt %. At 45 wt %, the permeability obtained by the poroelastic model decreases, and the Young's modulus increases. We suggest that macro (rather than micro) structural changes lead to this behavior due to the creation of a network of denser zones of chains at 45 wt % glucose. Our study paves the way for the design of calcium-alginate hydrogels with controlled structure for food and pharmaceutical applications in which interactions with glucose are of relevance.

Side chains affect the melt processing and stretchability of arabinoxylan biomass-based thermoplastic films.

Hydrophobization of hemicellulose causes melt processing and makes them stretchable thermoplastics. Understanding how native and/or appended side chains in various hemicelluloses after chemical modification affect melt processing and material properties can help in the development of products for film packaging and substrates for stretchable electronics applications. Herein, we describe a one-step and two-step strategy for the fabrication of flexible and stretchable thermoplastics prepared by compression molding of two structurally different arabinoxylans (AX). For one-step synthesis, the n-butyl glycidyl ether epoxide ring was opened to the hydroxyl group, resulting in the introduction of alkoxide side chains. The first step in the two-step synthesis was periodate oxidation. Because the melt processability for AXs having low arabinose to xylose ratio (araf/xylp<0.5) have been limited, two structurally distinct AXs extracted from wheat bran (AXWB, araf/xylp = 3/4) and barley husk (AXBH, araf/xylp = 1/4) were used to investigate the effect of araf/xylp and hydrophobization on the melt processability and properties of the final material. Melt compression processability was achieved in AXBH derived samples. DSC and DMA confirmed that the thermoplastics derived from AXWB and AXBH had dual and single glass transition (Tg) characteristics, respectively, but the thermoplastics derived from AXBH had lower stretchability (maximum 160%) compared to the AXWB samples (maximum 300%). Higher araf/xylp values, and thus longer alkoxide side chains in AXWB-derived thermoplastics, explain the stretchability differences.

Fat tissue equivalent phantoms for microwave applications by reinforcing gelatin with nanocellulose.

Tissue mimicking phantom materials with thermal and dielectric equivalence are vital for the development of microwave diagnostics and treatment. The current phantoms representing fat tissue are challenged by mechanical integrity at relevant temperatures coupled with complex production protocols. We have employed two types of nanocellulose (cellulose nanocrystals and oxidized cellulose nanocrystals) as reinforcement in gelatin stabilized emulsions for mimicking fat tissue. The nanocellulose-gelatin stabilized emulsions were evaluated for their dielectric properties, the moduli-temperature dependence using small deformation rheology, stress-strain behavior using large deformation, and their compliance to quality assurance guidelines for superficial hyperthermia. All emulsions had low permittivity and conductivity within the lower microwave frequency band, accompanied by fat equivalent thermal properties. Small deformation rheology showed reduced temperature dependence of the moduli upon addition of nanocellulose, independent of type. The cellulose nanocrystals gelatin reinforced emulsion complied with the quality assurance guidelines. Hence, we demonstrate that the addition of cellulose nanocrystals to gelatin stabilized emulsions has the potential to be used as fat phantoms for the development of microwave diagnostics and treatment.

Hydrophobization of arabinoxylan with n-butyl glycidyl ether yields stretchable thermoplastic materials.

Hemicelluloses are regarded as one of the first candidates for the development of value-added materials due to their renewability, abundance, and functionality. However, because most hemicelluloses are brittle, they can only be processed as a solution and cannot be processed using industrial melt-based polymer processing techniques. In this study, arabinoxylan (AX) was hydrophobized by incorporating butyl glycidyl ether (BuGE) into the hydroxyl groups through the opening of the BuGE epoxide ring, yielding alkoxy alcohols with terminal ethers. The formed BuGE derivatives were melt processable and can be manufactured into stretchable thermoplastic films through compression molding, which has never been done before with hemicellulose modified in a single step. The structural and thermomechanical properties of the one-step synthesis approach were compared to those of a two-step synthesis with a pre-activation step to demonstrate its robustness. The strain at break for the one-step synthesized AX thermoplastic with 3 mol of BuGE is ≈200%. These findings suggest that thermoplastic polymers can be composited with hemicelluloses or that thermoplastic polymers made entirely of hemicelluloses can be designed as packaging and stretchable electronics supports.

Oxidized xylan additive for nanocellulose films - A swelling modifier.

Polymeric wood hemicelluloses are depicted to join cellulose, starch and chitosan as key polysaccharides for sustainable materials engineering. However, the approaches to incorporate hemicelluloses in emerging bio-based products are challenged by lack of specific benefit, other than the biomass-origin, although their utilization would contribute to sustainable material use since they currently are a side stream that is not valorized. Here we demonstrate wood-xylans as swelling modifiers for neutral and charged nanocellulose films that have already entered the sustainable packaging applications, however, suffer from humidity sensitivity. The oxidative modification is used to modulate the water-solubility of xylan and hence enable adsorption in an aqueous environment. A high molecular weight grade, hence less water-soluble, adsorbed preferentially on the neutral surface while the adsorbed amount on a negatively charged surface was independent of the molecular weight, and hence, solubility. The adsorption of the oxidized xylans on a neutral cellulose surface resulted in an increase in the amount of water in the film while on the negatively charged cellulose the total amount of water decreased. The finding of synergy of two hygroscopic materials to decrease swelling in hydrophilic bio-polymer films demonstrates the oxidized macromolecule xylan as structurally functional component in emerging cellulose products.

Maximizing the oil content in polysaccharide-based emulsion gels for the development of tissue mimicking phantoms.

Formulations based on agar and κ-carrageenan were investigated for the production of emulsion gels applicable as tissue mimicking phantoms. The effects of the polysaccharide matrix, the oil content and the presence of surfactants on the micro-/nanostructure, rheology, and mechanical and dielectric properties were investigated. Results showed a high capacity of the agar to stabilize oil droplets, producing gels with smaller (10-21 µm) and more uniform oil droplets. The addition of surfactants allowed increasing the oil content and reduced the gel strength and stiffness down to 57 % and 34 %, respectively. The permittivity and conductivity of the gels were reduced by increasing the oil content, especially in the agar gels (18.8 and 0.05 S/m, respectively), producing materials with dielectric properties similar to those of low-water content tissues. These results evidence the suitability of these polysaccharides to design a variety of tissue mimicking phantoms with a broad range of mechanical and dielectric properties.

Polyglutamine expanded Ataxin-7 induces DNA damage and alters FUS localization and function.

Polyglutamine (polyQ) diseases, such as Spinocerebellar ataxia type 7 (SCA7), are caused by expansions of polyQ repeats in disease specific proteins. The sequestration of vital proteins into aggregates formed by polyQ proteins is believed to be a common pathological mechanism in these disorders. The RNA-binding protein FUS has been observed in polyQ aggregates, though if disruption of this protein plays a role in the neuronal dysfunction in SCA7 or other polyQ diseases remains unclear. We therefore analysed FUS localisation and function in a stable inducible PC12 cell model expressing the SCA7 polyQ protein ATXN7. We found that there was a high degree of FUS sequestration, which was associated with a more cytoplasmic FUS localisation, as well as a decreased expression of FUS regulated mRNAs. In contrast, the role of FUS in the formation of γH2AX positive DNA damage foci was unaffected. In fact, a statistical increase in the number of γH2AX foci, as well as an increased trend of single and double strand DNA breaks, detected by comet assay, could be observed in mutant ATXN7 cells. These results were further corroborated by a clear trend towards increased DNA damage in SCA7 patient fibroblasts. Our findings suggest that both alterations in the RNA regulatory functions of FUS, and increased DNA damage, may contribute to the pathology of SCA7.

Oxidation Level and Glycidyl Ether Structure Determine Thermal Processability and Thermomechanical Properties of Arabinoxylan-Derived Thermoplastics.

Developing flexible, stretchable, and thermally processable materials for packaging and stretchable electronic applications from polysaccharide-based polymers contributes to the smooth transition of the fossil-based economy to the circular bioeconomy. We present arabinoxylan (AX)-based thermoplastics obtained by ring-opening oxidation and subsequent reduction (dA-AX) combined with hydrophobization with three different glycidyl ethers [n-butyl (BuGE), isopropyl (iPrGE), and 2-ethyl hexyl (EtHGE) glycidyl ether]. We also investigate the relationship between structural composition, thermal processing, and thermomechanical properties. BuGE- and iPrGE-etherified dA-AXs showed glass-transition temperatures (Tg) far below their degradation temperatures and gave thermoplastic materials when compression-molded at 140 °C. The BuGE (3 mol)-etherified dA-AX films at 19 and 31% oxidation levels show 244% (±42) and 267% (±72) elongation, respectively. In contrast, iPrGE-dA-AX samples with shorter and branched terminals in the side chains had a maximum of 60% (±19) elongation. No studies have reported such superior elongation of AX thermoplastic films and its relationship with molar substitution and Tg. These findings have implications on the strategic development of chemical modification routes using commercial polymer processing technologies and on fine-tuning structures and properties when specific polysaccharide-based polymers are used to engineer bio-based products for film, packaging, and substrates for stretchable electronic applications.

Advanced structural characterisation of agar-based hydrogels: Rheological and small angle scattering studies.

Agar-based extracts from Gelidium sesquipedale were generated by heat and combined heat-sonication, with and without the application of alkali pre-treatment. Pre-treatment yielded extracts with greater agar contents; however, it produced partial degradation of the agar, reducing its molecular weight. Sonication produced extracts with lower agar contents and decreased molecular weights. A gelation mechanism is proposed based on the rheological and small angle scattering characterization of the extracts. The formation of strong hydrogels upon cooling was caused by the association of agarose chains into double helices and bundles, the sizes of which depended on the agar purity and molecular weight. These different arrangements at the molecular scale consequently affected the mechanical performance of the obtained hydrogels. Heating of the hydrogels produced a gradual disruption of the bundles; weaker or smaller bundles were formed upon subsequent cooling, suggesting that the process was not completely reversible.