Filament Stiffness Research Articles

Personalised 3D Printed Medicines: Optimising Material Properties for Successful Passive Diffusion Loading of Filaments for Fused Deposition Modelling of Solid Dosage Forms.

Although not readily accessible yet to many community and hospital pharmacists, fuse deposition modelling (FDM) is a 3D printing technique that can be used to create a 3D pharmaceutical dosage form by employing drug loaded filaments extruded via a nozzle, melted and deposited layer by layer. FDM requires printable filaments, which are commonly manufactured by hot melt extrusion, and identifying a suitable extrudable drug-excipient mixture can sometimes be challenging. We propose here the use of passive diffusion as an accessible loading method for filaments that can be printed using FDM technology to allow for the fabrication of oral personalised medicines in clinical settings. Utilising Hansen Solubility Parameters (HSP) and the concept of HSP distances (Ra) between drug, solvent, and filament, we have developed a facile pre-screening tool for the selection of the optimal combination that can provide a high drug loading (a high solvent-drug Ra, >10, and an intermediate solvent–filament Ra value, ~10). We have identified that other parameters such as surface roughness and stiffness also play a key role in enhancing passive diffusion of the drug into the filaments. A predictive model for drug loading was developed based on Support Vector Machine (SVM) regression and indicated a strong correlation between both Ra and filament stiffness and the diffusion capacity of a model BCS Class II drug, nifedipine (NFD), into the filaments. A drug loading, close to 3% w/w, was achieved. 3D printed tablets prepared using a PVA-derived filament (Hydrosupport, 3D Fuel) showed promising characteristics in terms of dissolution (with a sustained release over 24 h) and predicted chemical stability (>3 years at 25 °C/60% relative humidity), similar to commercially available NFD oral dosage forms. We believe FDM coupled with passive diffusion could be implemented easily in clinical settings for the manufacture of tailored personalised medicines, which can be stored over long periods of time (similar to industrially manufactured solid dosage forms).

Mutual Sliding Motion of Wrapped Filaments for Biomedical and Engineering Applications.

Here we aim at understanding and modeling of macroscopic interactions and sliding motion of curved filaments during muscles' isometric action in which tension is developed without overall contraction. A generic dynamic model of a curved elastic filament undergoing sliding, twisting, and unraveling around a cylindrical filament affected by the interfilament friction force is developed in full detail. In particular, the dynamic equations describing the general sliding motion of a curved filament wrapped around a cylindrical filament and pulled by a constant force applied to a free end are derived and solved numerically; the other end of the curved filament is considered to be fixed at the cylindrical one. The model predicts propagation of an elastic wave over the wrapped filament determined by the filament stiffness and the interfilament friction. The wrapped filament deformation and its ultimate arrest are predicted, and the final configurations of such filaments are revealed. Accordingly, the wrapped filament strain is predicted as a function of time for different values of the friction coefficient. The potential applications and possible biomechanical links of the proposed generic model are also discussed.

Interfacial Stereocomplexation To Strengthen Fused Deposition Modeled Poly(lactide) Welds

The interlayer stiffness of fused deposition modeled (FDM) parts is significantly impaired by the slow diffusion and re-entangling of polymer chains across filament interfacial welds in the build direction. To mechanically reinforce FDM interfaces, an approach based on timing and spatial direction of stereocomplexation across weld interfaces is introduced via alternating deposition of enantiomerically opposite poly(lactides). Fundamental insight into the kinetics and spatial distribution of homo- and stereocrystallization at FDM interfaces was successfully reconstructed in 2 and 3 dimensions by micrometer resolved Fourier transform infrared microscopy and synchrotron wide-angle X-ray diffraction tomography. The rate of isothermal stereocomplexation and consequential interfacial stiffening increases with decreased absolute and relative molar masses. The spatial distribution of stereocrystals is governed by the relative molar masses and the extent of interdiffusion, under not only isothermal but also nonisothermal FDM conditions. The net local heat dosage, which depends on print speed, governs the length scales of stereocomplexation and thus mechanical reinforcement. Interfacial stereocomplexation of poly(lactides) in FDM leads to a distinct 40% increase in stiffness and nucleation of bulk filaments, aiding in thermodynamic and geometrical stability.

Molecular structure influence on bending behavior of polyester filaments examined using a modified cantilever bending test

ABSTRACTAn experimental setup used to measure the bending stiffness of polymer filaments based on existing cantilever approaches is presented. Additionally, it is utilized to evaluate the influence of molar mass and long‐chain branching on filaments of poly(butylene terephthalate), modified with a multifunctional chain extender. Using shear rheological analysis process limits for extrusion of the modified material were revealed, indicated by a change of the curve shape of van Gurp–Palmen plots. For the processable modified materials, it was found that increasing molar mass and degree of long‐chain branching, caused by the modification, raise the bending stiffness of filaments. Also the Young's modulus was found to increase with the amount of chain extender used, while no difference could be generated by using different linear molar grades of the polymer. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 47952.

Buckling soft tensegrities: Fickle elasticity and configurational switching in living cells

Tensegrity structures are special architectures made by floating compressed struts kept together by a continuous system of tensioned cables. Their existence in a mechanically stable form is decided by the possibility of finding geometrical configurations such that pre-stressed tendons and bars can ensure self-equilibrium of the forces transmitted through the elastic network, the overall stiffness of which finally depends on both the rigidity of the compressed elements and the cables’ pre-stress. The multiplicity of shapes that tensegrity structures can assume and their intrinsic capability to be deployable and assembled, so storing (and releasing) elastic energy, have motivated their success as paradigm –pioneeringly proposed three decades ago by the intuition of Donald E. Ingber– to explain some underlying mechanisms regulating dynamics of living cells. The interlaced structure of the cell cytoskeleton, constituted by actin microfilaments, intermediate filaments and microtubules which continuously change their spatial organization and pre-stresses through polymerization/depolymerization processes, seems in fact to steer migration, adhesion and cell division by obeying the tensegrity construct. Even though rough calculations lead to estimate discrepancies of less than one order of magnitude when comparing axial stiffness of actin filaments (cables) and microtubules (struts) and recent works have shown bent microtubules among stretched filaments, no one has yet tried to remove the standard hypothesis of rigid struts in tensegrity structures when used to idealize the cell cytoskeleton mechanical response. With reference to the 30-element tensegrity cell paradigm, we thus introduce both compressibility and bendability of the struts and accordingly rewrite the theory to simultaneously take into account geometrical non-linearity (i.e. large deformations) and hyper-elasticity of both tendons and bars, so abandoning the classical linear stress-strain constitutive assumptions. By relaxing the hypothesis of rigidity of the struts, we demonstrate that some quantitative confirmations and many related extreme and somehow counter-intuitive mechanical behaviors actually exploited by cells for storing/releasing energy, resisting to applied loads and deforming by modulating their overall elasticity and shape through pre-stress changes and instability-guided configurational switching, can be all theoretically found. It is felt that the proposed new soft-strut tensegrity model could pave the way for a wider use of engineering models in cell mechanobiology and in designing bio-inspired materials and soft robots.

Nebulin increases thin filament stiffness and force per cross-bridge in slow-twitch soleus muscle fibers.

Nebulin (Neb) is associated with the thin filament in skeletal muscle cells, but its functions are not well understood. For this goal, we study skinned slow-twitch soleus muscle fibers from wild-type (Neb+) and conditional Neb knockout (Neb-) mice. We characterize cross-bridge (CB) kinetics and the elementary steps of the CB cycle by sinusoidal analysis during full Ca2+ activation and observe that Neb increases active tension 1.9-fold, active stiffness 2.7-fold, and rigor stiffness 3.0-fold. The ratio of stiffness during activation and rigor states is 62% in Neb+ fibers and 68% in Neb- fibers. These are approximately proportionate to the number of strongly attached CBs during activation. Because the thin filament length is 15% shorter in Neb- fibers than in Neb+ fibers, the increase in force per CB in the presence of Neb is ∼1.5 fold. The equilibrium constant of the CB detachment step (K 2), its rate (k 2), and the rate of the reverse force generation step (k -4) are larger in Neb+ fibers than in Neb- fibers. The rates of the force generation step (k 4) and the reversal detachment step (k -2) change in the opposite direction. These effects can be explained by Le Chatelier's principle: Increased CB strain promotes less force-generating state(s) and/or detached state(s). Further, when CB distributions among the six states are calculated, there is no significant difference in the number of strongly attached CBs between fibers with and without Neb. These results demonstrate that Neb increases force per CB. We also confirm that force is generated by isomerization of actomyosin (AM) from the AM.ADP.Pi state (ADP, adenosine diphophate; Pi, phosphate) to the AM*ADP.Pi state, where the same force is maintained after Pi release to result in the AM*ADP state. We propose that Neb changes the actin (and myosin) conformation for better ionic and hydrophobic/stereospecific AM interaction, and that the effect of Neb is similar to that of tropomyosin.

Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle

Nebulin is a giant sarcomeric protein that spans along the actin filament in skeletal muscle, from the Z-disk to near the thin filament pointed end. Mutations in nebulin cause muscle weakness in nemaline myopathy patients, suggesting that nebulin plays important roles in force generation, yet little is known about nebulin's influence on thin filament structure and function. Here, we used small-angle X-ray diffraction and compared intact muscle deficient in nebulin (using a conditional nebulin-knockout, Neb cKO) with control (Ctrl) muscle. When muscles were activated, the spacing of the actin subunit repeat (27 Å) increased in both genotypes; when converted to thin filament stiffness, the obtained value was 30 pN/nm in Ctrl muscle and 10 pN/nm in Neb cKO muscle; that is, the thin filament was approximately threefold stiffer when nebulin was present. In contrast, the thick filament stiffness was not different between the genotypes. A significantly shorter left-handed (59 Å) thin filament helical pitch was found in passive and contracting Neb cKO muscles, as well as impaired tropomyosin and troponin movement. Additionally, a reduced myosin mass transfer toward the thin filament in contracting Neb cKO muscle was found, suggesting reduced cross-bridge interaction. We conclude that nebulin is critically important for physiological force levels, as it greatly stiffens the skeletal muscle thin filament and contributes to thin filament activation and cross-bridge recruitment.

Self-reinforced biobased composites based on high stiffness PLA yarns

In self-reinforced polymer composites (SRPC) the same polymer material forms both the reinforcing fibre and the matrix phase. The current project aims to develop a biobased alternative for these composites using polylactic acid (PLA). Both the development of the reinforcing fibres, the modelling and production of the composites are being studied. The stiffness of the PLA filaments could be increased to 9 GPa by optimising processing parameters during extrusion. After consolidation to composites, promising results are obtained showing that the stiffness of the PLA SRPC can match the requirements of currently used commercial self-reinforced polypropylene composites (ca 4GPa).

Dry brushing: Does it improve plaque removal? A secondary analysis.

This article is a secondary analysis comparing the effects on plaque removal of brushing with a dry toothbrush and brushing with a prewetted toothbrush. The participants had been included in two previous experiments involving single-brushing exercises. The 46 non-dental participants were systemically healthy and ≥18years of age. In the first experiment, the control intervention was brushing with a prewetted toothbrush, while during the second experiment it was brushing with a dry toothbrush. Both experiments scored plaque before and after the brushing exercises and assessed participants' perception. The data of these two previous experiments were compared in this secondary analysis. Plaque score reduction following brushing with a dry toothbrush was 58%, while with a prewetted toothbrush, it was 57%. The mean plaque index score reduction of 0.08 between a dry and a prewetted toothbrush was not significant (P=.096). Prewetting the participants' toothbrush had no influence on the perception of toothbrush filament stiffness (P=.410) nor on the perception of cleaning capability (P=.449). In both experiments, brushing without dentifrice was judged to be unpleasant. On average, following a 2-minute brushing exercise, plaque scored were reduced by 57% or more. Dry brushing did not contribute significantly to toothbrush efficacy. The participants did not find that prewetting a toothbrush influenced the cleaning capability and filament stiffness.

Label-free Imaging and Bending Analysis of Microtubules by ROCS Microscopy and Optical Trapping

Mechanical manipulation of single cytoskeleton filaments and their monitoring over long times is difficult because of fluorescence bleaching or phototoxic protein degradation. The integration of label-free microscopy techniques, capable of imaging freely diffusing, weak scatterers such as microtubules (MTs) in real-time, and independent of their orientation, with optical trapping and tracking systems, would allow many new applications. Here, we show that rotating-coherent-scattering microscopy (ROCS) in dark-field mode can also provide strong contrast for structures far from the coverslip such as arrangements of isolated MTs and networks. We could acquire thousands of images over up to 30 min without loss in image contrast or visible photodamage. We further demonstrate the combination of ROCS imaging with fast and nanometer-precise 3D interferometric back-focal-plane tracking of multiple beads in time-shared optical traps using acoustooptic deflectors to specifically construct and microrheologically probe small microtubule networks with well-defined geometries. Thereby, we explore the frequency-dependent elastic response of single microtubule filaments between 0.5 Hz and 5 kHz, which allows for investigating their viscoelastic response up to the fourth-order bending mode. Our spectral analysis reveals constant filament stiffness at low frequencies and frequency-dependent stiffening following a power law ∼ωp with a length-dependent exponent p(L). We find further evidence for the dependence of the MT persistence length on the contour length L, which is still controversially debated. We could also demonstrate slower stiffening at high frequencies for longer filaments, which we believe is determined by the molecular architecture of the MT. Our results shed new light on the nanomechanics of this essential, multifunctional cytoskeletal element and pose new questions about the adaptability of the cytoskeleton.

Interaction between toothpaste abrasivity and toothbrush filament stiffness on the development of erosive/abrasive lesions in vitro

To investigate the loss of enamel and dentin surface caused by the interaction between abrasives in toothpaste and toothbrush filament stiffness. The study followed a 2 (high-level or low-level abrasive; silica) × 3 (filament stiffness; soft, medium or hard) × 2 (cycling time; 3 or 5 days) factorial design. Polished bovine enamel and dentin specimens (n = 8 each per group) were subjected to 5 days of erosion/abrasion cycling: erosion (5 minutes, four times daily, 0.3% citric acid, pH 3.75); abrasion (15 seconds, twice daily, 45 strokes each, 150 g load, automated brushing machine); and fluoride treatment [15 seconds with abrasion and 45 seconds without abrasion; 275 p.p.m. fluoride (F- ) as sodium fluoride (NaF) in abrasive slurry]. Enamel and dentin specimens were exposed to artificial saliva between erosion and abrasion/F- treatment (1 hour) and at all other times (overnight). Non-contact profilometry was used to determine surface loss (SL) after 3 and 5 days of cycling. Data were analysed using three-way analysis of variance (ANOVA) (factors: abrasive/filament stiffness/time), with separate analyses conducted for enamel and dentin. For enamel, only 'cycling time' was found to affect SL, with 5 days of cycling resulting in a greater SL than 3 days of cycling. Overall, there was little SL for enamel (range: 0.76-1.85 μm). For dentin (SL range: 1.87-5.91 μm), significantly higher SL was found for 5 days of cycling versus 3 days of cycling, with particularly large differences for hard stiffness/high-level abrasive and medium stiffness/low-level abrasive. For high-level abrasive, after 5 days of cycling hard stiffness resulted in significantly higher SL than did medium stiffness, with no other significant differences according to stiffness. Overall, high-level abrasive resulted in significantly higher SL than did low-level abrasive, with strong effects for all combinations, except medium stiffness after 5 days. The interplay between abrasivity and filament stiffness appears to be more relevant for dentin than for enamel.

DNA Nanotubes as a Versatile Tool to Study Semiflexible Polymers.

Mechanical properties of complex, polymer-based soft matter, such as cells or biopolymer networks, can be understood in neither the classical frame of flexible polymers nor of rigid rods. Underlying filaments remain outstretched due to their non-vanishing backbone stiffness, which is quantified via the persistence length (lp), but they are also subject to strong thermal fluctuations. Their finite bending stiffness leads to unique, non-trivial collective mechanics of bulk networks, enabling the formation of stable scaffolds at low volume fractions while providing large mesh sizes. This underlying principle is prevalent in nature (e.g., in cells or tissues), minimizing the high molecular content and thereby facilitating diffusive or active transport. Due to their biological implications and potential technological applications in biocompatible hydrogels, semiflexible polymers have been subject to considerable study. However, comprehensible investigations remained challenging since they relied on natural polymers, such as actin filaments, which are not freely tunable. Despite these limitations and due to the lack of synthetic, mechanically tunable, and semiflexible polymers, actin filaments were established as the common model system. A major limitation is that the central quantity lp cannot be freely tuned to study its impact on macroscopic bulk structures. This limitation was resolved by employing structurally programmable DNA nanotubes, enabling controlled alteration of the filament stiffness. They are formed through tile-based designs, where a discrete set of partially complementary strands hybridize in a ring structure with a discrete circumference. These rings feature sticky ends, enabling the effective polymerization into filaments several microns in length, and display similar polymerization kinetics as natural biopolymers. Due to their programmable mechanics, these tubes are versatile, novel tools to study the impact of lp on the single-molecule as well as the bulk scale. In contrast to actin filaments, they remain stable over weeks, without notable degeneration, and their handling is comparably straightforward.

The relationship between dentine hypersensitivity, dietary acid intake and erosive tooth wear

ObjectivesTo assess the interplay between dietary acid intake, toothbrushing and erosive tooth wear as aetiological factors in self-reported DH Materials and methodsThis is a secondary analysis on previously collected data from 600 participants with (n=300) and without (n=300) severe erosive tooth wear. Participants recruited from restorative clinics of King’s College London Dental Institute (REC Ref 14/EM/1171) were questioned on their self-reported DH, frequency and timing of dietary acid intake, habits associated with consumption of dietary acids and tooth brushing habits. Erosive tooth wear was assessed using Basic Erosive Wear Examination (BEWE). Differences in diet and brushing habits between those with self-reported DH and those without were analysed using descriptives and logistic regression in SPSS vers. 24. ResultsOf those reporting with DH (n=272), a greater number spent ≥10min eating fruit per sitting, (n=46) and had a drinking habit such as sipping, swishing or holding drinks in the mouth (n=72) compared to those without DH (n=26 OR 2.72, 95%CI [1.32–5.61], p=0.007 and n=38 OR2.33, 95%CI [1.40–3.88], p=0.001, respectively). More DH participants used a soft toothbrush (n=36) compared to those without DH (n=18 OR 2.35, 95%CI [1.20–4.59], p=0.013). No association was observed with frequency of daily toothbrushing or dietary acid intake. ConclusionContact time between the tooth and the acid may be a more important risk factor for DH compared to frequency of dietary acid intake or frequency of toothbrushing. Other possible aetiological factors should be considered. Clinicaltrials.gov numberNCT02449434 Clinical significanceIncreased contact time with dietary acids and sipping swishing or holding drinks in the mouth prior to swallowing should be addressed as an aetiological factor in DH. Toothpaste abrasivity and toothbrush filament stiffness may play a greater role in DH compared to frequency of toothbrushing.

How geometric frustration shapes twisted fibres, inside and out: competing morphologies of chiral filament assembly.

Chirality frustrates and shapes the assembly of flexible filaments in rope-like, twisted bundles and fibres by introducing gradients of both filament shape (i.e. curvature) and packing throughout the structure. Previous models of chiral filament bundle formation have shown that this frustration gives rise to several distinct morphological responses, including self-limiting bundle widths, anisotropic domain (tape-like) formation and topological defects in the lateral inter-filament order. In this paper, we employ a combination of continuum elasticity theory and discrete filament bundle simulations to explore how these distinct morphological responses compete in the broader phase diagram of chiral filament assembly. We show that the most generic model of bundle formation exhibits at least four classes of equilibrium structure-finite-width, twisted bundles with isotropic and anisotropic shapes, with and without topological defects, as well as bulk phases of untwisted, columnar assembly (i.e. 'frustration escape'). These competing equilibrium morphologies are selected by only a relatively small number of parameters describing filament assembly: bundle surface energy, preferred chiral twist and stiffness of chiral filament interactions, and mechanical stiffness of filaments and their lateral interactions. Discrete filament bundle simulations test and verify continuum theory predictions for dependence of bundle structure (shape, size and packing defects of two-dimensional cross section) on these key parameters.

Modeling the Actin.myosin ATPase Cross-Bridge Cycle for Skeletal and Cardiac Muscle Myosin Isoforms

Modeling the complete actin.myosin ATPase cycle has always been limited by the lack of experimental data concerning key steps of the cycle, because these steps can only be defined at very low ionic strength. Here, using human β-cardiac myosin-S1, we combine published data from transient and steady-state kinetics to model a minimal eight-state ATPase cycle. The model illustrates the occupancy of each intermediate around the cycle and how the occupancy is altered by changes in actin concentration for [actin] = 1–20Km. The cycle can be used to predict the maximal velocity of contraction (by motility assay or sarcomeric shortening) at different actin concentrations (which is consistent with experimental velocity data) and predict the effect of a 5 pN load on a single motor. The same exercise was repeated for human α-cardiac myosin S1 and rabbit fast skeletal muscle S1. The data illustrates how the motor domain properties can alter the ATPase cycle and hence the occupancy of the key states in the cycle. These in turn alter the predicted mechanical response of the myosin independent of other factors present in a sarcomere, such as filament stiffness and regulatory proteins. We also explore the potential of this modeling approach for the study of mutations in human β-cardiac myosin using the hypertrophic myopathy mutation R453C. Our modeling, using the transient kinetic data, predicts mechanical properties of the motor that are compatible with the single-molecule study. The modeling approach may therefore be of wide use for predicting the properties of myosin mutations.

Measurement of the Persistence Length of Cytoskeletal Filaments using Curvature Distributions

Cytoskeletal filaments such as microtubules and actin filaments play important roles in the mechanical integrity of cells and the ability of cells to respond to their environment. Measuring the mechanical prop- erties of cytoskeletal structures is crucial for gaining insight into intracellular mechanical stresses and their role in regulating cellular processes. One of the ways to characterize these mechanical properties is by measuring their persistence length, the average length over which filaments stay straight. There are several approaches in the literature for measuring filament deformations, including Fourier analysis of images obtained using fluorescence microscopy. Here, we show how curvature distributions can be used as an alternative tool to quantify bio-filament deformations, and investigate how the apparent stiffness of filaments depends on the resolution and noise of the imaging system. We present analytical calculations of the scaling curvature distributions as a function of filament discretization, and test our predictions by comparing Monte Carlo simulations to results from existing techniques. We also apply our approach to microtubules and actin filaments obtained from in vitro gliding assay experiments with high densities of non-functional motors, and calculate the persistence length of these filaments. The presented curvature analysis is significantly more accurate compared to existing approaches for small data sets, and can be readily applied to both in vitro or in vivo filament data through the use of an ImageJ plugin we provide.

筋芽細胞の分化の初期段階における細胞表面硬さとアクチン線維の変化

筋芽細胞の分化の初期段階における細胞表面硬さとアクチン線維の変化

A multiscale biomechanical model of platelets: Correlating with in-vitro results

Using dissipative particle dynamics (DPD) combined with coarse grained molecular dynamics (CGMD) approaches, we developed a multiscale deformable platelet model to accurately describe the molecular-scale intra-platelet constituents and biomechanical properties of platelets in blood flow. Our model includes the platelet bilayer membrane, cytoplasm and an elaborate elastic cytoskeleton. Correlating numerical simulations with published in-vitro experiments, we validated the biorheology of the cytoplasm, the elastic response of membrane to external stresses, and the stiffness of the cytoskeleton actin filaments, resulting in an accurate representation of the molecular-level biomechanical microstructures of platelets. This enabled us to study the mechanotransduction process of the hemodynamic stresses acting onto the platelet membrane and transmitted to these intracellular constituents. The platelets constituents continuously deform in response to the flow induced stresses. To the best of our knowledge, this is the first molecular-scale platelet model that can be used to accurately predict platelets activation mechanism leading to thrombus formation in prosthetic cardiovascular devices and in vascular disease processes. This model can be further employed to study the effects of novel therapeutic approaches of modulating platelet properties to enhance their shear resistance via mechanotransduction pathways.

Effects of Thermal Tension Transients on the Muscle Crossbridge

The transverse thermal fluctuations of the myosin molecule are significant. This paper explores the contribution of lateral myosin bending to the developed crossbridge force and power stroke. The equipartition theorem is used to calculate the mode amplitudes for myosin bending. Crossbridge axial force [Formula: see text] and power stroke [Formula: see text] are developed by transverse in-plane fluctuations along the [Formula: see text]- and [Formula: see text]-axes. Practical applications include the effects of temperature on the flexibility of the myosin molecule stiffness and tension, relevant to man-made fabrication of synthetic muscle using micromachines and nanowires. Scaling laws for the [Formula: see text] bending amplitude depend on filament length, mode number, and stiffness, as [Formula: see text], and (EI)[Formula: see text]. This paper quantifies the effects of thermal motion on the mechanics of miniature molecular motors, including the muscle crossbridge.

Cations Stiffen Actin Filaments by Adhering a Key Structural Element to Adjacent Subunits.

Ions regulate the assembly and mechanical properties of actin filaments. Recent work using structural bioinformatics and site-specific mutagenesis favors the existence of two discrete and specific divalent cation binding sites on actin filaments, positioned in the long axis between actin subunits. Cation binding at one site drives polymerization, while the other modulates filament stiffness and plays a role in filament severing by the regulatory protein, cofilin. Existing structural methods have not been able to resolve filament-associated cations, and so in this work we turn to molecular dynamics simulations to suggest a candidate binding pocket geometry for each site and to elucidate the mechanism by which occupancy of the “stiffness site” affects filament mechanical properties. Incorporating a magnesium ion in the “polymerization site” does not seem to require any large-scale change to an actin subunit’s conformation. Binding of a magnesium ion in the “stiffness site” adheres the actin DNase-binding loop (D-loop) to its long-axis neighbor, which increases the filament torsional stiffness and bending persistence length. Our analysis shows that bound D-loops occupy a smaller region of accessible conformational space. Cation occupancy buries key conserved residues of the D-loop, restricting accessibility to regulatory proteins and enzymes that target these amino acids.