An analytical approach of filament bundle swinging dynamics, Part II: identifying equivalent dynamic constitutive parameters of filament bundles

A filament bundle is a type of yarn, which is composed of nearly parallel and highly oriented polymer monofilaments. Due to its nonlinearity both in material constitutive properties and structure, the filament bundle possesses nonlinear viscoelastic properties. It is important to study the dynamic behavior of the filament bundle accurately during its high-speed movement. Therefore, an accurate expression of the constitutive relation of the filament bundle is an essential prerequisite for its dynamic simulation and analysis. Continued the previous study in Part I: modeling filament bundle method, in this paper, an approach was proposed to identify the equivalent dynamic constitutive parameters of the filament bundle considering frequency-dependent characteristics. Firstly, the identification formulas of the dynamic elastic modulus and viscoelastic coefficients were derived based on the Kelvin model. Then, a testing method of the cross-sectional parameters of the filament bundle under a certain tension was proposed, and the testing device was developed to obtain the area of the filament bundle; The dynamic loading test of the bundle filament was conducted in a DMA Q800 dynamic mechanical tester. Thirdly, the equivalent dynamic elastic modulus and viscoelastic coefficients were obtained through the experimental test. Finally, an analytical method was proposed to verify the correctness of experimental results through simulation. The results show that the excitation frequency has a significant influence on the dynamic elastic modulus and viscoelastic coefficient, and the curves of the equivalent dynamic elastic modulus and viscoelastic coefficient present nonlinear variation characteristics.

Cryo-EM structures of CTP synthase filaments reveal mechanism of pH-sensitive assembly during budding yeast starvation

Many metabolic enzymes self-assemble into micron-scale filaments to organize and regulate metabolism. The appearance of these assemblies often coincides with large metabolic changes as in development, cancer, and stress. Yeast undergo cytoplasmic acidification upon starvation, triggering the assembly of many metabolic enzymes into filaments. However, it is unclear how these filaments assemble at the molecular level and what their role is in the yeast starvation response. CTP Synthase (CTPS) assembles into metabolic filaments across many species. Here, we characterize in vitro polymerization and investigate in vivo consequences of CTPS assembly in yeast. Cryo-EM structures reveal a pH-sensitive assembly mechanism and highly ordered filament bundles that stabilize an inactive state of the enzyme, features unique to yeast CTPS. Disruption of filaments in cells with non-assembly or pH-insensitive mutations decreases growth rate, reflecting the importance of regulated CTPS filament assembly in homeotstasis.

Characterization of heterologously expressed fibril filaments, a shape and motility determining cytoskeletal protein of the helical bacterium Spiroplasma

Fibril is a constitutive filament forming cytoskeletal protein of unidentified fold, exclusive to members of genus Spiroplasma. It is hypothesized to undergo conformational changes necessary to bring about Spiroplasma motility through changes in body helicity. However, in the absence of a cofactor such as nucleotide that binds to the protein and drives polymerization, the mechanism driving conformational changes in fibril remains unknown. Sodium dodecyl sulphate (SDS) solubilized the fibril filaments and facilitated fibril purification by affinity chromatography. An alternate protocol for obtaining enriched insoluble fibril filaments has been standardized using density gradient centrifugation method. Visualization of purified protein using electron microscopy demonstrated that it forms filament bundles. Probable domain boundaries of fibril protein were identified based on mass spectrometric analysis of proteolytic fragments. Presence of both α-helical and β-sheet signatures in FT-IR measurements suggests that fibril filaments consist of assembly of folded globular domains, and not a β-strand based aggregation similar to amyloid fibrils.

INF2-mediated actin filament reorganization confers intrinsic resilience to neuronal ischemic injury

During early stages of ischemic brain injury, glutamate receptor hyperactivation mediates neuronal death via osmotic cell swelling. Here we show that ischemia and excess NMDA receptor activation, conditions that trigger neuronal swelling, cause actin filaments to undergo a rapid and extensive reorganization within the somatodendritic compartment. Normally, Factin is concentrated within dendritic spines, with relatively little Factin in the dendrite shaft. However, beginning <5 min after incubation of neurons with NMDA, Factin depolymerizes within dendritic spines and polymerizes into long, stable filament bundles within the dendrite shaft and soma. A similar actinification of the somatodendritic compartment occurs after oxygen/glucose deprivation in vitro, and in mouse brain after photothrombotic stroke in vivo. Following transient, sub-lethal NMDA exposure these actin changes spontaneously reverse within 1-2 hours. A combination of Na+, Cl-, water, and Ca2+ entry are all necessary, but not individually sufficient, for induction of actinification. Spine F-actin depolymerization is also required. Actinification is driven by activation of the Factin polymerization factor inverted formin2 (INF2). Silencing of INF2 renders neurons more vulnerable to NMDA induced membrane leakage and cell death, and formin inhibition markedly increases ischemic infarct severity in vivo. These results show that ischemia induced actin filament reorganization within the dendritic compartment is an intrinsic pro-survival response that protects neurons from death induced by swelling.

Experimental Study on Angular Flexural Performance of Multiaxis Three Dimensional (3D) Polymeric Carbon Fiber/Cementitious Concretes

Multiaxis three-dimensional (3D) continuous polymeric carbon fiber/cementitious concretes were introduced. Their angular (off-axis) flexural properties were experimentally studied. It was found that the placement of the continuous carbon fibers and their in-plane angular orientations in the pristine concrete noticeably influenced the angular flexural strength and the energy absorption behavior of the multiaxis 3D concrete composite. The off-axis flexural strength of the uniaxial (C-1D-(0°)), biaxial (C-2D-(0°), and C-2D-(90°)), and multiaxial (C-4D-(0°), C-4D-(+45°) and C-4D-(−45°)) concrete composites were outstandingly higher (from 36.84 to 272.43%) than the neat concrete. Their energy absorption capacities were superior compared to the neat concrete. Fractured four directional polymeric carbon fiber/cementitious matrix concretes limited brittle matrix failure and a broom-like fracture phenomenon on the filament bundles, filament-matrix debonding and splitting, and minor filament entanglement. Multiaxis 3D polymeric carbon fiber concrete, especially the C-4D structure, controlled the crack phenomena and was considered a damage-tolerant material compared to the neat concrete.

Cryo-EM Structures of CTP Synthase Filaments Reveal Mechanism of pH-Sensitive Assembly During Budding Yeast Starvation

ABSTRACTMany metabolic enzymes self-assemble into micron-scale filaments to organize and regulate metabolism. The appearance of these assemblies often coincides with large metabolic changes as in development, cancer, and stress. Yeast undergo cytoplasmic acidification upon starvation, triggering the assembly of many metabolic enzymes into filaments. However, it is unclear how these filaments assemble at the molecular level and what their role is in the yeast starvation response. CTP Synthase (CTPS) assembles into metabolic filaments across many species. Here, we characterize in vitro polymerization and investigate in vivo consequences of CTPS assembly in yeast. Cryo-EM structures reveal a pH-sensitive assembly mechanism and highly ordered filament bundles that stabilize an inactive state of the enzyme, features unique to yeast CTPS. Disruption of filaments in cells with non-assembly or hyper-assembly mutations decreases growth rate, reflecting the importance of regulated CTPS filament assembly in homeotstasis.

Braiding Dynamics in Semiflexible Filament Bundles under Oscillatory Forcing

We examine the nonequilibrium production of topological defects—braids—in semiflexible filament bundles under cycles of compression and tension. During these cycles, the period of compression facilitates the thermally activated pair production of braid/anti-braid pairs, which then may separate when the bundle is under tension. As a result, appropriately tuned alternating periods of compression and extension should lead to the proliferation of braid defects in a bundle so that the linear density of these pairs far exceeds that expected in the thermal equilibrium. Secondly, we examine the slow extension of braided bundles under tension, showing that their end-to-end length creeps nonmonotonically under a fixed force due to braid deformation and the motion of the braid pair along the bundle. We conclude with a few speculations regarding experiments on semiflexible filament bundles and their networks.

Epithelial sodium channel (ENaC) in GtoPdb v.2021.2

The epithelial sodium channels (ENaC) are located on the apical membrane of epithelial cells in the kidney tubules, lung, respiratory tract, male and female reproductive tracts, sweat and salivary glands, placenta, colon, and some other organs [9, 13, 22, 21, 42]. In these epithelia, Na+ ions flow from the extracellular fluid into the cytoplasm of epithelial cells via ENaC. The Na+ ions are then pumped out of the cytoplasm into the interstitial fluid by the Na+/K+ ATPase located on the basolateral membrane [36]. As Na+ is one of the major electrolytes in the extracellular fluid (ECF), osmolarity change initiated by the Na+ flow is accompanied by a flow of water accompanying Na+ ions [6]. Thus, ENaC has a central role in regulating ECF volume and blood pressure, primarily via its function in the kidney [37]. The expression of ENaC subunits, hence its activity, is regulated by the renin-angiotensin-aldosterone system, and other factors involved in electrolyte homeostasis [37, 30]. In the respiratory tract and female reproductive tract, large segments of the epithelia are composed of multi-ciliated cells. In these cells, ENaC is located along the entire length of the cilia that cover the cell surface [15]. Cilial location greatly increases ENaC density per cell surface and allows ENaC to serve as a sensitive regulator of osmolarity of the periciliary fluid throughout the whole depth of the fluid bathing the cilia [15]. In contrast to ENaC, CFTR (ion transporter defective in cystic fibrosis) is located on non-cilial cell-surface [15]. In the vas deferens segment of the male reproductive tract, the luminal surface is covered by microvilli and stereocilia projections with backbones composed of actin filament bundles [42]. In these cells, both ENaC and the water channel aquaporin AQP9 are localized on these projections and also in the basal and smooth muscle layers [42]. Thus, ENaC function is also essential for the clearance of respiratory airways, transport of germ cells, fertilization, implantation, and cell migration [15, 22].