Resistance of the fiber-derived geotextile from Typha domingensis submitted to field degradation.

Geotextiles made from plant fibers creates a suitable environment for plant growth as part of soil bioengineering techniques. The faster decomposition of plant fiber geotextiles compared to synthetic ones demands the use of composites that enhance their waterproofing and extend their durability in the environment. The objective of this work was to evaluate the resistance of a geotextile made with Thypha domingensis to degradation caused by climatic variables. Tensile strength tests were conducted in the laboratory in order to evaluate the degradation of geotextiles treated with single and double layers of waterproofing resin. Based on Scanning Electron Microscopy (SEM) images, it was verified that applying double layer of waterproofing resin delays the fibers degradation up to 120 days of exposure to the effects of climatic variables other than temperature. The maximum resistance losses due to the geotextile's exposure to degradation were statistically significant for all three treatments: control-without waterproofing resin, with one layer resin, and with two layers resin. Therefore, waterproofing resin, provides a long-term protective solution for geotextiles made from cattail fibers.

The Molecular Mechanism of the Response of Rice to Arsenic Stress and Effective Strategies to Reduce the Accumulation of Arsenic in Grain.

Rice (Oryza sativa L.) is the staple food for more than 50% of the world's population. Owing to its growth characteristics, rice has more than 10-fold the ability to enrich the carcinogen arsenic (As) than other crops, which seriously affects world food security. The consumption of rice is one of the primary ways for humans to intake As, and it endangers human health. Effective measures to control As pollution need to be studied and promoted. Currently, there have been many studies on reducing the accumulation of As in rice. They are generally divided into agronomic practices and biotechnological approaches, but simultaneously, the problem of using the same measures to obtain the opposite results may be due to the different species of As or soil environments. There is a lack of systematic discussion on measures to reduce As in rice based on its mechanism of action. Therefore, an in-depth understanding of the molecular mechanism of the accumulation of As in rice could result in accurate measures to reduce the content of As based on local conditions. Different species of As have different toxicity and metabolic pathways. This review comprehensively summarizes and reviews the molecular mechanisms of toxicity, absorption, transport and redistribution of different species of As in rice in recent years, and the agronomic measures to effectively reduce the accumulation of As in rice and the genetic resources that can be used to breed for rice that only accumulates low levels of As. The goal of this review is to provide theoretical support for the prevention and control of As pollution in rice, facilitate the creation of new types of germplasm aiming to develop without arsenic accumulation or within an acceptable limit to prevent the health consequences associated with heavy metal As as described here.

Chasing high and stable wheat grain mineral content: Mining diverse spring genotypes under induced drought stress.

Climate change-induced drought has an effect on the nutritional quality of wheat. Here, the impact of drought at different plant stages on mineral content in mature wheat was evaluated in 30 spring-wheat lines of diverse backgrounds (modern, old and wheat-rye-introgressions). Genotypes with rye chromosome 3R introgression showed a high accumulation of several important minerals, including Zn and Fe, and these also showed stability across drought conditions. High Se content was found in genotypes with chromosome 1R. Old cultivars (K, Mg, Na, P and S) and 2R introgression lines (Fe, Ca, Mn, Mg and Na) demonstrated high mineral yield at early and late drought, respectively. Based on the low nutritional value often reported for modern wheat and negative climate effects on the stability of mineral content and yield, genes conferring high Zn/Fe, Se, and stable mineral yield under drought at various plant stages should be explicitly explored among 3R, 1R, old and 2R genotypes, respectively.

Molecular and Systems Biology Approaches for Harnessing the Symbiotic Interaction in Mycorrhizal Symbiosis for Grain and Oil Crop Cultivation.

Mycorrhizal symbiosis, the mutually beneficial association between plants and fungi, has gained significant attention in recent years due to its widespread significance in agricultural productivity. Specifically, arbuscular mycorrhizal fungi (AMF) provide a range of benefits to grain and oil crops, including improved nutrient uptake, growth, and resistance to (a)biotic stressors. Harnessing this symbiotic interaction using molecular and systems biology approaches presents promising opportunities for sustainable and economically-viable agricultural practices. Research in this area aims to identify and manipulate specific genes and pathways involved in the symbiotic interaction, leading to improved cereal and oilseed crop yields and nutrient acquisition. This review provides an overview of the research frontier on utilizing molecular and systems biology approaches for harnessing the symbiotic interaction in mycorrhizal symbiosis for grain and oil crop cultivation. Moreover, we address the mechanistic insights and molecular determinants underpinning this exchange. We conclude with an overview of current efforts to harness mycorrhizal diversity to improve cereal and oilseed health through systems biology.

Flours from fermented lentil and quinoa grains as ingredients with new techno-functional properties.

The need to provide novel, nutritious plant-based products requires seeking high-value, sustainable protein sources, like quinoa and lentils, having an increased digestibility and lacking antinutrients. Fungal fermentation has evidenced enhanced nutritional value of flours obtained from these grains. However, research into techno-functional properties, essential to the new product development, is lacking. This study investigated the techno-functional properties of flours made from lentil and quinoa after fermenting them with Pleurotus ostreatus and subjecting them to two drying techniques (lyophilisation and hot air drying). In both cases, the fermentation led to noteworthy improvements in swelling and water holding capacity, especially in those lyophilised than those dried. In contrast, the emulsifying, foaming, thickening, and gelling capacities decreased significantly. The loss of abilities was more severe for dried grains than for lyophilized ones. The thermomechanical analysis of the fermented flours showed lower thickening and gelling potential compared to untreated flours. Microscopy images revealed that the state and structure of starch granules were affected by both fermentation and drying processes. Starch granules in lentils were partly pre-gelatinised and trapped in the cotyledon cell, resulting in limited thickening and gelling abilities. In contrast, in quinoa, starch underwent pre-gelatinisation and retrogradation during the fermentation process, promoting the production of resistant starch and increasing fibre content. This study presents the potential of treated flours as ingredients possessing unique attributes compared to protein and fibre-rich conventional products.

An analysis of lncRNAs related to fiber quality and the discovery of their target genes in a Gossypium hirsutum line with Gossypium mustelinum introgression.

KEY MESSAGE: Analysis of fiber quality lncRNAs and their target genes from a pair of Gossypium mustelinum near-isogenic lines provide new prospects for improving the fiber quality of Upland cotton. Long noncoding RNAs (lncRNAs) are an important part of genome transcription and play roles in a wide range of biological processes in plants. In this research, a pair of near-isogenic cotton lines, namely, a Gossypium mustelinum introgression line (IL9) with outstanding fiber quality and its recurrent Upland cotton parent (PD94042), were used as the experimental materials. Cotton fibers were selected for lncRNA sequencing at 17 and 21 days post-anthesis. A total of 2693 differentially expressed genes were identified. In total, 5841 lncRNAs were ultimately screened, from which 163 differentially expressed lncRNAs were identified. Target genes of the lncRNAs were predicted by two different methods: cis and trans. Some of the target genes were related to cell components, membrane components, plant hormone signal transduction and catalytic metabolism, and the results indicated that there might also be important effects on the development of fiber. Four differentially expressed target genes related to fiber quality (Gomus.D05G015100, Gomus.A05G281300, Gomus.A12G023400 and Gomus.A10G226800) were screened through gene function annotation, and the functions of these four genes were verified through virus-induced gene silencing (VIGS). Compared to the negative controls, plants in which any of these four genes were silenced showed significant reductions in fiber strength. In addition, the plants in which the Gomus.A12G023400 gene was silenced showed a significant reduction in fiber uniformity, whereas the plants in which Gomus.A05G281300 was silenced showed a significant increase in fiber fineness as measured via micronaire. Our results showed that these genes play different roles during fiber development, impacting fiber quality.

Conformational properties of heterogeneous arabinoxylan protein gums from corn bran and distillers grains in comparison with gum arabic.

The molecular structure and conformation of arabinoxylan-protein gum, commonly referred as corn fiber gum (CFG) were analyzed by high-performance size-exclusion chromatography (HPSEC) coupled with RI, UV, light scattering and viscometer detectors. CFG had a heterogeneous structure. The detailed conformation of CFG at different molecular weights was compared with that of hemicellulose fiber gum (HFG) from dried distiller's grains with solubles and gum arabic. The CFG molecules mainly had random coil conformation; only 10 % of them exhibited rigid rod conformation. Approximately 80 % of the CFG had a molecular weight between 105 and 105.4 Da, while the other 20 % of molecules were between 105.4 and 1.5 × 107.7 Da. The overall conformational properties of CFG and HFG were closer but differed from that of gum arabic. The intrinsic viscosity and radius of gyration of both CFG and HFG were greater than those of gum arabic although the average molecular weight of CFG and HFG was lower. The protein and carbohydrate were covalently linked in CFG molecules as shown by the HPSEC-multiple detectors combined with partial acid hydrolysis. Based on the detailed conformation of CFG and the methylation analysis, 1D and 2D NMR spectroscopy results, the molecular structure of CFG was proposed.

Phenotypic and Proteomic Insights into Differential Cadmium Accumulation in Maize Kernels.

The contamination of agricultural soil with cadmium (Cd), a heavy metal, poses a significant environmental challenge, affecting crop growth, development, and human health. Previous studies have established the pivotal role of the ZmHMA3 gene, a P-type ATPase heavy metal transporter, in determining variable Cd accumulation in maize grains among 513 inbred lines. To decipher the molecular mechanism underlying mutation-induced phenotypic differences mediated by ZmHMA3, we conducted a quantitative tandem mass tag (TMT)-based proteomic analysis of immature maize kernels. This analysis aimed to identify differentially expressed proteins (DEPs) in wild-type B73 and ZmHMA3 null mutant under Cd stress. The findings demonstrated that ZmHMA3 accumulated higher levels of Cd compared to B73 when exposed to varying Cd concentrations in the soil. In comparison to soil with a low Cd concentration, B73 and ZmHMA3 exhibited 75 and 142 DEPs, respectively, with 24 common DEPs shared between them. ZmHMA3 showed a higher induction of upregulated genes related to Cd stress than B73. Amino sugar and nucleotide sugar metabolism was specifically enriched in B73, while phenylpropanoid biosynthesis, nitrogen metabolism, and glyoxylate and dicarboxylate metabolism appeared to play a more significant role in ZmHMA3. This study provides proteomics insights into unraveling the molecular mechanism underlying the differences in Cd accumulation in maize kernels.

Sequential Extraction and Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy Monitoring in the Biorefining of Brewer's Spent Grain.

The brewing industry plays a significant role in producing a substantial annual volume of by-products, which contributes to the global accumulation of food waste. The primary by-product generated is brewer's spent grain (BSG), a lignocellulosic biomass rich in proteins, fiber, and moisture content. Leveraging biorefining and valorization techniques for BSG represents a promising strategy to enhance sustainability, resilience, and circularity within the brewing chain. To date, most studies have focused on extracting proteins from BSG. Yet, it is crucial to note that the fiber part of BSG also holds considerable potential for biorefining processes. This study introduces a novel sequential extraction method designed to integrally recover the major components of BSG. Notably, it introduces a reactive extraction approach that enables the simultaneous extraction and tuneable functionalization of the hemicellulose component. Additionally, the study assesses the utility of the attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy as a user-friendly tool to monitor and evaluate the effectiveness of the fractionation process. This spectroscopic technique can provide valuable insights into the changes and composition of BSG throughout the extraction process.

Recent Progress in Distiller's Grains: Chemical Compositions and Biological Activities.

Distiller's grains (DGs) are solid mixtures that remain after the production of alcoholic beverages. A large amount of DGs is produced each year during the brewing process. Currently, they are mostly used as a feedstock or substrate in the feed industry. However, the lack of a comprehensive understanding of the chemical composition of DGs is a major constraint on their further development and application for high-value-added usages. Some studies were published on the bioactive constituents of DGs in several different types of journals. Data were therefore collated to provide a comprehensive overview of these natural products. DGs are rich in phenols, phytosterols, and fatty acids, in addition to general lipid and protein constituents. These compounds and their related extracts possess diverse biological activities, including antioxidant, anti-inflammatory, and anti-hyperglycaemic effects. We hope that this review will provide research incentives for the further development and utilisation of DGs to develop high-value-added products.

Effects of straw return with potassium fertilizer on the stem lodging resistance, grain quality and yield of spring maize (Zea mays L.).

This experiment aimed to study the effects of straw return combined with potassium fertilizer on stem lodging resistance, grain quality, and yield of spring maize. The objective was to provide a scientific basis for the rational utilization of Inner Mongolia spring maize straw and potassium fertilizer resources. The test material used was 'Xianyu 335', and the study was conducted in three ecological regions from east to west of Inner Mongolia (Tumochuan Plain Irrigation Area, Hetao Plain Irrigation Area, and Lingnan Warm Dry Zone). A split-plot design was employed, with the straw return method as the main plot and potassium fertilizer dosage as the secondary plot. We determined the stem resistance index, grain quality, and yield. The results showed that both straw return and potassium application improved stem lodging resistance, grain quality, and maize yield. Combining straw return with the reasonable application of potassium fertilizer enhanced the effectiveness of potassium fertilizer, increased lodging resistance, maize yield, and improved grain quality and yield stability. Under the straw return treatment, with potassium application compared to no potassium application, significant increases were observed in maize plant height, stem diameter, dry weight of stems, stem compressive strength, stem bending strength, grain protein content, yield, straw potassium accumulation content, and soil available potassium content. These increases were up to 30.79 cm, 2.63 mm, 15.40 g, 74.93 N/mm2, 99.65 N/mm2, 13.68%, 3142.43 kg/hm2, 57.97 kg/hm2, and 19.80 mg/kg, respectively. Therefore, the interaction of straw return and potassium fertilizer was found to be the most effective measure for maintaining high-yield and stress-resistant cultivation, improving grain quality, and optimizing the management of straw and potassium fertilizer resources. This approach is suitable for promotion and application in the spring maize growing areas of Inner Mongolia.

Assessment of biodegradation of lignocellulosic fiber-based composites - A systematic review.

Lignocellulosic fiber-reinforced polymer composites are the most extensively used modern-day materials with low density and better specific strength specifically developed to render better physical, mechanical, and thermal properties. Synthetic fiber-reinforced composites face some serious issues like low biodegradability, non-environmentally friendly, and low disposability. Lignocellulosic or natural fiber-reinforced composites, which are developed from various plant-based fibers and animal-based fibers are considered potential substitutes for synthetic fiber composites because they are characterized by lightweight, better biodegradability, and are available at low cost. It is very much essential to study end-of-life (EoL) conditions like biodegradability for the biocomposites which occur commonly after their service life. During biodegradation, the physicochemical arrangement of the natural fibers, the environmental conditions, and the microbial populations, to which the natural fiber composites are exposed, play the most influential factors. The current review focuses on a comprehensive discussion of the standards and assessment methods of biodegradation in aerobic and anaerobic conditions on a laboratory scale. This review is expected to serve the materialists and technologists who work on the EoL behaviour of various materials, particularly in natural fiber-reinforced polymer composites to apply these standards and test methods to various classes of biocomposites for developing sustainable materials.

Mechanical superiority of Pseudoxytenanthera bamboo for sustainable engineering solutions.

The advancement in natural fibre composites has replaced synthetic fibres in various commercial sectors. Bamboo species possess high mechanical properties due to their lignocellulosic fibre content, which makes them suitable for engineering applications and potential alternatives to solid wood. However, despite Bamboo being composed of 130 genera and 1700 different species, out of which many still remains underexplored. In this study, we investigated the, Lignocellulosic profiling, fibre strength, and mechanical characterization of two species of Pseudoxytenanthera Bamboo: Pseudoxytenanthera ritchiei, Pseudopxytenanthera stocksii, and the results obtained were compared with Bambusa balcooa, one of the priority species of bamboo identified by The International Plant Genetic Resources Institute (IPGRI). BET (Brunauer-Emmett-Teller) was used to quantify the samples' density, while SEM-EDX and FTIR spectroscopy were used for elemental analysis. The samples were then subjected to tensile test in addition, thermogravimetric analysis and water absorption test were carried out for the three species. The results showed that Pseudoxytenanthera species possessed superior chemical and mechanical characteristics compared to the priority species of bamboo used for composites. Out of the two Pseudoxytenanthera species studied, Pseudoxytenanthera stocksii exhibited the highest values of cellulose, hemicellulose, lignin, pectin, ash, carbon, and silicon, indicating its chemical superiority. Moreover, Pseudoxytenanthera stocksii also showed higher mechanical values for tensile strength, making it suitable for a variety of engineering applications. The TGA values also indicated that Pseudoxytenanthera stocksii is stable at high temperatures when compared with other natural fibres.

Extraction and characterization of natural cellulosic fiber from Mariscus ligularis plant as potential reinforcement in composites.

In this paper, fiber from the Mariscus ligularis (ML) plant was extracted and investigated as a naturally derived fiber for its potential as a reinforcement material for composite applications. Physical, morphological, chemical, thermal, and mechanical property analyses of the Mariscus ligularis fiber (MLF) were performed to evaluate its suitability as a reinforcement material while also generating useful data to serve as the basis for its selection in the development of new composite materials. Physical and morphological analysis results showed MLF as a lightweight fiber of diameter 243.6 µm and density 768.59 kg/m3 with a very rough surface that provides excellent interfacial bonding performance. Chemical and thermal results show MLF has mainly cellulose as its crystallized phase, with cellulose and wax contents of 58.32 % and 0.73 %, respectively, and possesses a 72.23 % crystallinity index and a 3.15 nm crystallite size with thermal stability up to 258 °C. The mechanical results show that the tensile strength, elastic modulus, strain to failure, and microfibril angle were in the ranges of 109-134 MPa, 3.27-5.06 GPa, 3.32-9.13 %, and 13.35-20.33°, respectively. These findings show MLF as a potential reinforcement material.

Fibres and Colorectal Cancer: Clinical and Molecular Evidence.

Colorectal cancer (CRC) is one of the leading causes of mortality for cancer in industrialized countries. The link between diet and CRC is well-known, and presumably CRC is the type of cancer which is most influenced by dietary habits. In Western countries, an inadequate dietary intake of fibers is endemic, and this could be a driving factor in the increase of CRC incidence. Indeed, several epidemiologic studies have elucidated an inverse relationship between daily fiber intake and risk of CRC. Long-term prognosis in CRC survivors is also dependent on dietary fibers. Several pathogenetic mechanisms may be hypothesized. Fibers may interfere with the metabolism of bile acids, which may promote colon carcinogenesis. Further, fibers are often contained in vegetables which, in turn, contain large amounts of antioxidant agents like resveratrol, polyphenols, or phytoestrogens. Moreover, fibers can be digested by commensal flora, thus producing compounds such as butyrate, which exerts an antiproliferative effect. Finally, fibers may modulate gut microbiota, whose composition has shown to be associated with CRC onset. In this regard, dietary interventions based on high-fiber-containing diets are ongoing to prevent CRC development, especially in patients with high potential for this type of tumor. Despite the fact that outcomes are preliminary, encouraging results have been observed.

A review of recent developments in kenaf fiber/polylactic acid composites research.

Kenaf fiber has recently garnered exponential interest as reinforcement in composite materials across diverse industries owing to its superior mechanical attributes, ease of manufacture, and inherent biodegradability. In the discourse of this review, various methods of manufacturing kenaf/Polylactic acid (PLA) composites have been discussed meticulously, as delineated in recently published scientific literatures. This paper delves into the chemical modification of kenaf fiber, examining its consequential impact on tensile strength and thermal stability of the kenaf/PLA composites. Further, this review illuminates the role of innovative 3D printing techniques and fiber orientation in augmenting the mechanical robustness of the kenaf/PLA composites. Simultaneously, recent insightful explorations into the acoustic properties of the kenaf/PLA composites, underscoring their potential as sustainable alternative to conventional materials have been reviewed. Serving as a comprehensive repository of knowledge, this review paper holds immense value for researchers aiming to utilize the capabilities of kenaf fiber reinforced PLA composites.

Primary care services in the English NHS: are they a thorn in the side of integrated care systems? A qualitative analysis.

BACKGROUND: As integrated care systems are embedded across England there are regions where the integration process has been evaluated and continues to evolve. Evaluation of these integrated systems contributes to our understanding of the challenges and facilitators to this ongoing process. This can support integrated care systems nationwide as they continue to develop. We describe how two integrated care partnerships in different localities, at differing stages of integration with contrasting approaches experienced challenges specifically when integrating with primary care services. The aim of this analysis was to focus on primary care services and how their existing structures impacted on the development of integrated care systems. METHODS: We carried out an exploratory approach to re-analysing our previously conducted 51 interviews as part of our prior evaluations of integrated health and care services which included primary care services. The interview data were thematically analysed, focussing on the role and engagement of primary care services with the integrated care systems in these two localities. RESULTS: Four key themes from the data are discussed: (i) Workforce engagement (engagement with integration), (ii) Organisational communication (information sharing), (iii) Financial issues, (iv) Managerial information systems (data sharing, IT systems and quality improvement data). We report on the challenges of ensuring the workforce feel engaged and informed. Communication is a factor in workforce relationships and trust which impacts on the success of integrated working. Financial issues highlight the conflict between budget decisions made by the integrated care systems when primary care services are set up as individual businesses. The incompatibility of information technology systems hinders integration of care systems with primary care. CONCLUSIONS: Integrated care systems are national policy. Their alignment with primary care services, long considered to be the cornerstone of the NHS, is more crucial than ever. The two localities we evaluated as integration developed both described different challenges and facilitators between primary care and integrated care systems. Differences between the two localities allow us to explore where progress has been made and why.

A Hands-On Guide to Generate Spatial Gene Expression Profiles by Integrating scRNA-seq and 3D-Reconstructed Microscope-Based Plant Structures.

Transcriptome profiles of individual cells in the plant are strongly dependent on their relative position. Cell differentiation is associated with tissue-specific transcriptomic changes. For that reason, it is important to study gene expression changes in a spatial context, and therefore to link those to potential morphological changes over developmental time. Even though great experimental advances have been made in recording spatial gene expression profiles, those attempts are limited in the plant field. New computational approaches attempt to solve this problem by integrating spatial expression profiles of few marker genes with single-cell/single-nuclei RNA-seq (scRNA-seq) methodologies. In this chapter, we provide a practical guide on how to predict gene expression patterns in a 3D plant structure by combining scRNA-seq data and 3D microscope-based reconstructed expression profiles of a small set of reference genes. We also show how to visualize these results.

Honey bees (Apis mellifera) modify plant-pollinator network structure, but do not alter wild species' interactions.

Honey bees (Apis mellifera) are widely used for honey production and crop pollination, raising concern for wild pollinators, as honey bees may compete with wild pollinators for floral resources. The first sign of competition, before changes appear in wild pollinator abundance or diversity, may be changes to wild pollinator interactions with plants. Such changes for a community can be measured by looking at changes to metrics of resource use overlap in plant-pollinator interaction networks. Studies of honey bee effects on plant-pollinator networks have usually not distinguished whether honey bees alter wild pollinator interactions, or if they merely alter total network structure by adding their own interactions. To test this question, we experimentally introduced honey bees to a Canadian grassland and measured plant-pollinator interactions at varying distances from the introduced hives. We found that honey bees increased the network metrics of pollinator and plant functional complementarity and decreased interaction evenness. However, in networks constructed from just wild pollinator interactions, honey bee abundance did not affect any of the metrics calculated. Thus, all network structural changes to the full network (including honey bee interactions) were due only to honey bee-plant interactions, and not to honey bees causing changes in wild pollinator-plant interactions. Given widespread and increasing use of honey bees, it is important to establish whether they affect wild pollinator communities. Our results suggest that honey bees did not alter wild pollinator foraging patterns in this system, even in a year that was drier than the 20-year average.

Programmable spatial deformation by controllable off-center freestanding 4D printing of continuous fiber reinforced liquid crystal elastomer composites.

Owing to their high deformation ability, 4D printed structures have various applications in origami structures, soft robotics and deployable mechanisms. As a material with programmable molecular chain orientation, liquid crystal elastomer is expected to produce the freestanding, bearable and deformable three-dimensional structure. However, majority of the existing 4D printing methods for liquid crystal elastomers can only fabricate planar structures, which limits their deformation designability and bearing capacity. Here we propose a direct ink writing based 4D printing method for freestanding continuous fiber reinforced composites. Continuous fibers can support freestanding structures during the printing process and improve the mechanical property and deformation ability of 4D printed structures. In this paper, the integration of 4D printed structures with fully impregnated composite interfaces, programmable deformation ability and high bearing capacity are realized by adjusting the off-center distribution of the fibers, and the printed liquid crystal composite can carry a load of up to 2805 times its own weight and achieve a bending deformation curvature of 0.33 mm-1 at 150 °C. This research is expected to open new avenues for creating soft robotics, mechanical metamaterials and artificial muscles.