Electric-field induced modulation of amorphous protein aggregates: polarization, deformation, and reorientation.

Proteins in their native state are only marginally stable and tend to aggregate. However, protein misfolding and condensation are often associated with undesired processes, such as pathogenesis, or unwanted properties, such as reduced biological activity, immunogenicity, or uncontrolled materials properties. Therefore, controlling protein aggregation is very important, but still a major challenge in various fields, including medicine, pharmacology, food processing, and materials science. Here, flexible, amorphous, micron-sized protein aggregates composed of lysozyme molecules reduced by dithiothreitol are used as a model system. The preformed amorphous protein aggregates are exposed to a weak alternating current electric field. Their field response is followed in situ by time-resolved polarized optical microscopy, revealing field-induced deformation, reorientation and enhanced polarization as well as the disintegration of large clusters of aggregates. Small-angle dynamic light scattering was applied to probe the collective microscopic dynamics of amorphous aggregate suspensions. Field-enhanced local oscillations of the intensity auto-correlation function are observed and related to two distinguishable elastic moduli. Our results validate the prospects of electric fields for controlling protein aggregation processes.

Direct visualization of local activities of long DNA strands via image-time correlation.

Bacteriophages with long DNA genomes are of interest due to their diverse mutations dependent on environmental factors. By lowering the ionic strength of a hydrophobic (PPh4Cl) antagonistic salt (at 1 mM), single long T4 DNA strand fluctuations were clearly observed, while condensed states of T4 DNA globules were formed above 5-10 mM salt. These long DNA strands were treated with fluorescently labeled probes, for which photo bleaching is often unavoidable over a short time of measurement. In addition, long (few tens of [Formula: see text]) length scales are required to have larger fields of view for better sampling, with shorter temporal resolutions. Thus, an optimization between length and time is crucial to obtain useful information. To facilitate the challenge of detecting large biomacromolecules, we here introduce an effective method of live image data analysis for direct visualization and quantification of local thermal fluctuations. The motions of various conformations for the motile long DNA strands were examined for the single- and multi-T4 DNA strands. We find that the unique correlation functions exhibit a relatively high-frequency oscillatory behavior superimposed on the overall slower decay of the correlation function with a splitting of amplitudes deriving from local activities of the long DNA strands. This work shows not only the usefulness of an image-time correlation for analyzing large biomacromolecules, but also provides insight into the effects of a hydrophobic antagonistic salt on active T4 bacteriophage long DNA strands, including thermal translocations in their electrostatic interactions.

Solvent-dependent morphology and anisotropic microscopic dynamics of cellulose nanocrystals under electric fields.

Cellulose nanocrystals (CNCs) are interesting for the construction of biomaterials for energy delivery and packaging purposes. The corresponding processing of CNCs can be optimized through the variation of intercellulose interactions by employing different types of solvents, and thereby varying the degree of cellulose hydrogen bonding. The aim of this work is (i) to show how different types of solvents affect the self-assembled morphology of CNCs, (ii) to study the microscopic dynamics and averaged orientations on the CNCs in aqueous suspensions, including the effect of externally imposed electric fields, and (iii) to explore the nonlinear optical response of CNCs. The homogeneity of self-assembled chiral-nematic phase depends on both the polarity of the solvent and the CNC concentration. The variation of the chiral-nematic pitch length with concentration, as determined from real-space and Fourier images, is found to be strongly solvent dependent. The anisotropic microdynamics of CNCs suspension exhibits two modes, related to diffusion parallel and perpendicular to the (chiral-) nematic director. We have found also the coupling between translational and orientational motion, due to existing correlation length of twisted nematic elasticity. Preliminary second-harmonic generation experiments are performed, which reveal that relatively high field strengths are required to reorient chiral-nematic domains of CNCs.

Equilibrium phase diagram and thermal responses of charged DNA-virus rod-suspensions at low ionic strengths.

The collective behavior of DNA is important for exploring new types of bacteria in the means of detection, which is greatly interested in the understanding of interactions between DNAs in living systems. How they self-organize themselves is a physical common phenomenon for broad ranges of thermodynamic systems. In this work, the equilibrium phase diagrams of charged chiral rods (fd viruses) at low ionic strengths (below a few mM) are provided to demonstrate both replicas of (or self-organized) twist orders and replica symmetry breaking near high concentration glass-states. By varying the ionic strengths, it appears that a critical ionic strength is obtained below 1-2 mM salt, where the twist and freezing of nematic domains diverge. Also, the microscopic relaxation is revealed by the ionic strength-dependent effective Debye screening length. At a fixed low ionic strength, the local orientations of twist are shown by two different length scales of optical pitch, in the chiral-nematic N* phase and the helical domains [Formula: see text], for low and high concentration, respectively. RSB occurs in several cases of crossing phase boundary lines in the equilibrium phase diagram of DNA-rod concentration and ionic strength, including long-time kinetic arrests in the presence of twist orders. The different pathways of PATH I, II and III are due to many-body effects of randomized orientations for charged fd rods undergoing long-range electrostatic interactions in bulk elastic medium. In addition, the thermal stability are shown for chiral pitches of the N* phase and the abnormal cooling process of a specific heat in a structural glass. Here, the concentration-driven twist-effects of charged DNA rods are explored using various experimental methods involving image-time correlation, microscopic dynamics in small angle dynamic light scattering, optical activity in second harmonic generation, and differential scanning calorimetry for the glass state.

Shockwave application enhances the effect of dentin desensitizer.

OBJECTIVES: The purpose of this study was to develop a new device that can improve the effect of desensitizer using shockwaves and to verify its efficacy. METHODS: A micro-shockwave generator was developed using a piezoelectric actuator (PIA-1000, piezosystem jena GmbH, Jena, Germany), an Arduino Uno microcontroller (Arduino, Torino, Italy), and a high voltage pulser (HVP-1000, piezosystem jena GmbH) at 700 V (400 A) and 100 µs. The occlusal surfaces of 20 extracted human upper and lower third molars without caries or restoration were reduced to expose the occlusal dentin, and the prepared occlusal surfaces were acid-etched with 32% phosphoric acid to remove the smear layer. The tooth specimens were connected to a fluid flow measurement instrument (nanoFlow, IB SYSTEMS, Seoul, Korea), permeability through dentin via dentinal fluid flow (DFF) was measured for 300 s, and the average DFF rate (Baseline DFF rate) was calculated. A desensitizer (SuperSeal, Phoenix Dental, Fenton, MI, USA) was applied to the acid-etched occlusal dentin surface of 10 randomly selected tooth specimens, left for 10 s, and rubbed with a microbrush for 30 s (Group 1). For the remaining teeth, the desensitizer was applied, and a shockwave (100 µm stroke, 10,000 G) was applied for 10 s (2 shots/s) and rubbed with a microbrush for 30 s (Group 2). After desensitizer application, subsequent DFF was measured for 600 s, and the average DFF rate was calculated (post-application DFF rate). DFF was continuously measured in real-time at 25 ±â€¯0.5 ℃ under a hydrostatic pressure of 25 cm. The percentage reduction in DFF rate after desensitizer application (with or without shockwave) was calculated with respect to baseline DFF rate. Data were analyzed with independent t-test (α = 0.05). RESULTS: For all tooth specimens, DFF rate decreased after desensitizer application irrespective of the presence of shockwaves. The percentage reduction in DFF rate of SuperSeal with shockwave (Group 2) was 42.8 ±â€¯19.0%, which was significantly higher than the 26.2 ±â€¯13.6% of the SuperSeal only group (Group 1) (p < 0.05). SIGNIFICANCE: Measurement of DFF change in real-time shows that shockwaves can help reduce dentin permeability beyond that SuperSeal dentin desensitizer produced alone.

Coupling of long-wavelength density fluctuations to orientations in cellulose nanocrystal suspensions under external fields.

Motivated by the development of cellulose-based functional materials, we investigate the microscopic dynamics of suspensions of cellulose nanocrystals (CNCs) at different ionic strengths, both in the absence and in the presence of AC electric fields and for various temperatures. A concentration of 5 wt % of the CNCs is chosen for which the dispersions are in the full chiral-nematic state at low ionic strengths. Dynamic light scattering is used to characterize the wave vector-dependent decay rates of number-density fluctuations. Contrary to an isotropic suspension, the dispersion relations (the wave vector dependence of the correlation-function decay rates) as obtained by means of depolarized light scattering are found to exhibit anomalous behavior. The dispersion relations, both without and with an external field, exhibit minima at small wave vectors, which is attributed to coupling of translational motion to the orientation of the CNCs, shown in the chiral-nematic state. The location of the minima is found to weakly depend on ionic strength and shifts significantly towards larger wave vectors upon applying an external electric field for sufficiently high ionic strengths. Finally, preliminary results are presented for smaller length-scale density fluctuations (at larger wave vectors) as a function of temperature, revealing the anisotropic mobilities in the chiral-nematic state of CNCs.

Anisotropic Diffusion and Phase Behavior of Cellulose Nanocrystal Suspensions.

In this paper, we use dynamic light scattering in polarized and depolarized modes to determine the translational and rotational diffusion coefficients of concentrated rodlike cellulose nanocrystals in aqueous suspension. Within the range of studied concentrations (1-5 wt %), the suspension starts a phase transition from an isotropic to an anisotropic state as shown by polarized light microscopy and viscosity measurements. Small-angle neutron scattering measurements also confirmed the start of cellulose nanocrystal alignment and a decreasing distance between the cellulose nanocrystals with increasing concentration. As expected, rotational and translational diffusion coefficients generally decreased with increasing concentration. However, the translational parallel diffusion coefficient was found to show a local maximum at the onset of the isotropic-to-nematic phase transition. This is attributed to the increased available space for rods to move along their longitudinal axis upon alignment. This increased parallel diffusion coefficient thus confirms the general idea that rodlike particles gain translational entropy upon alignment while paying the price for losing rotational degrees of freedom. Once the concentration increases further, diffusion becomes more hindered even in the aligned regions due to a reduction in the rod separation distance. This leads once again to a decrease in translational diffusion coefficients. Furthermore, the relaxation rate for fast mode translational diffusion (parallel to the long particle axis) exhibited two regimes of relaxation behavior at concentrations where significant alignment of the rods is measured. We attribute this unusual dispersive behavior to two length scales: one linked to the particle length (at large wavevector q) and the other to a twist fluctuation correlation length (at low wavevector q) along the cellulose nanocrystal rods that is of a larger length when compared to the actual length of rods and could be linked to the size of aligned domains.

Effect of Gelation on the Colloidal Deposition of Cellulose Nanocrystal Films.

One of the most important aspects in controlling colloidal deposition is manipulating the homogeneity of the deposit by avoiding the coffee-ring effect caused by capillary flow inside the droplet during drying. After our previous work where we achieved homogeneous deposition of cellulose nanocrystals (CNCs) from a colloidal suspension by reinforcing Marangoni flow over the internal capillary flow (Gençer et al. Langmuir 2017, 33 (1), 228-234), we now set out to reduce the importance of capillary flow inside a drying droplet by inducing gelation. In this paper, we discuss the effect of gelation on the deposition pattern and on the self-assembly of CNCs during droplet drying. CNC films were obtained by drop casting CNC suspensions containing NaCl and CaCl2 salts. A mixed methodology using rheological and depolarized dynamic light scattering was applied to understand the colloidal behavior of the CNCs. In addition, analysis of the mixture's surface tension, viscosity, and yield stress of the suspensions were used to gain deeper insights into the deposition process. Finally, the understanding of the gelation behavior in the drying droplet was used to exert control over the deposit where the coffee-ring deposit can be converted to a dome-shaped deposit.

Soft-mode of charged chiral fibrous viruses (fd).

The frictional forces in suspensions vary depending on the size, shape, and the surface of the particles, which are either charged or neutral. For anisotropic particles with no spatial gradient in the order parameter under external parameters, they exhibit either a continuous phase transition or "freezing" of the order parameter fluctuation. They are known as the collective soft-mode, which has a finite cutoff dispersion where the relaxation time diverges. From microscopic dynamics of charged chiral fd-viruses, the soft-mode is revealed with a rotation restoring "twist", obtained from both polarized (VV) and depolarized (VH) small angle dynamic light scattering. Here, I have found the minimum spatial coherence length at a lower I-N binodal concentration, which is due to the reverse of electrostatic repulsive forces with an increase in the concentration of charged chiral rods.

Flow instability due to coupling of shear-gradients with concentration: non-uniform flow of (hard-sphere) glasses.

Flow-induced instabilities that lead to non-uniform stationary flow profiles have been observed in many different soft-matter systems. Two types of instabilities that lead to banded stationary states have been identified, which are commonly referred to as gradient- and vorticity-banding. The molecular origin of these instabilities is reasonably well understood. A third type of instability that has been proposed phenomenologically [Europhys. Lett., 1986, 2, 129 and Phys. Rev. E, 1995, 52, 4009] is largely unexplored. Essential to this "Shear-gradient Concentration Coupling" (SCC-) instability is a mass flux that is induced by spatial gradients of the shear rate. A possible reason that this instability has essentially been ignored is that the molecular origin of the postulated mass flux is not clear, and no explicit expressions for the shear-rate and concentration dependence of the corresponding transport coefficient exist. It is therefore not yet known what types of flow velocity- and concentration-profiles this instability gives rise to. In this paper, an expression for the transport coefficient corresponding to the shear-gradient induced mass flux is derived in terms of the shear-rate dependent pair-correlation function, and Brownian dynamics simulations for hard-spheres are presented that specify the shear-rate and concentration dependence of the pair-correlation function. This allows to explicitly formulate the coupled advection-diffusion equation and an equation of motion for the suspension flow velocity. The inclusion of a non-local contribution to the stress turns out to be essential to describe the SCC-banding transition. The coupled equations of motion are solved numerically, and flow- and concentration-profiles are discussed. It is shown that the SCC-instability occurs within the glass state at sufficiently small shear rates, leading to a banded flow-profile where one of the bands is non-flowing.

An electric-field induced dynamical state in dispersions of charged colloidal rods.

The response of concentrated dispersions of charged colloids to low-frequency electric fields is governed by field-induced inter-colloidal interactions resulting from the polarization of electric double layers and the layer of condensed ions, association and dissociation of condensed ions, as well as hydrodynamic interactions through field-induced electro-osmotic flow. The phases and states that can be formed by such field-induced interactions are an essentially unexplored field of research. Experiments on concentrated suspensions of rod-like colloids (fd-virus particles), within the isotropic-nematic phase coexistence region, showed that a number of phases/states are induced, depending on the field amplitude and frequency [Soft Matter, 2010, 6, 273]. In particular, a dynamical state is found where nematic domains form and melt on a time scale of the order of seconds. We discuss the microscopic origin of this dynamical state, which is attributed to the cyclic, electric-field induced dissociation and association of condensed ions. A semi-quantitative theory is presented for the dynamics of melting and formation of nematic domains, including a model for the field-induced dissociation/association of condensed ions. The resulting equation of motion for the orientational order parameter is solved numerically for parameters complying with the fd-virus system. A limit-cycle is found, with a cycling-time that diverges at the transition line in the field-amplitude versus frequency plane where the dynamical state first appears, in accord with experimental findings.

Glass transition of repulsive charged rods (fd-viruses).

It has recently been shown that suspensions of long and thin charged fibrous viruses (fd) form a glass at low ionic strengths. The corresponding thick electric double layers give rise to long-ranged repulsive electrostatic interactions, which lead to caging and structural arrest at concentrations far above the isotropic-nematic coexistence region. Structural arrest and freezing of the orientational texture are found to occur at the same concentration. In addition, various types of orientational textures are equilibrated below the glass transition concentration, ranging from a chiral-nematic texture with a large pitch (of about 100 µm), an X-pattern, and a tightly packed domain texture, consisting of helical domains with a relatively small pitch (of about 10 µm) and twisted boundaries. The dynamics of both particles as well as the texture are discussed, below and above the glass transition. Dynamic light scattering correlation functions exhibit two dynamical modes, where the slow mode is attributed to the elasticity of helical domains. On approach of the glass-transition concentration, the slow mode increases in amplitude, while as the amplitudes of the fast and slow mode become equal at the glass transition. Finally, interesting features of the "transient" behaviors of charged fd-rod glass are shown as the initial caging due to structural arrest, the propagation of flow originating from stress release, and the transition to the final metastable glass state. In addition to the intensity correlation function, power spectra are presented as a function of the waiting time, at the zero-frequency limit that may access to the thermal anomalities in a charged system.

Spatio-temporal analysis of shrinkage vectors during photo-polymerization of composite.

OBJECTIVES: The purpose of this study was to validate a new method to investigate the polymerization shrinkage vectors of composite during light curing and to evaluate the overall utility and significance of the technique. METHODS: An optical instrument was developed to measure the location and direction of the polymerization shrinkage strain vectors of dental composite during light curing using a particle tracking method with computer vision. The measurement system consisted of a CCD color camera, a lens and a filter, and software for multi-particle tracking. A universal hybrid composite (Z250, 3M ESPE, St. Paul MN, USA) was molded into thin disk-shaped specimens (un-bonded and bonded) or filled into a cavity within a tooth slab (bonded). The composite surface was coated with fluorescent particles prior to light curing. The images of the fluorescent particles were stored at 2 frames/s for 10 min, and the movements of the particles on the composite surface were tracked with computer vision during curing. The polymerization shrinkage strain vectors as a function of time and location were analyzed. The volume shrinkage of the composite was also measured for comparison. RESULTS: The linear and volume shrinkage of the composite at 10 min were 0.75 (0.12)% and 2.26 (0.18)%, respectively. The polymerization shrinkage vectors were directed toward the center of the specimen and were isotropic in all directions when the composite was allowed to shrinkage freely without bonding. In contrast, the shrinkage vectors were directed toward the bonding surface and were anisotropic when the composite was bonded to a fixed wall. The regional displacement vectors of composite in a tooth cavity were dependent on the location, depth and time. SIGNIFICANCE: The new instrument was able to measure the regional linear shrinkage strain vectors over an entire surface of a composite specimen as a function of time and location. Therefore, this instrument can be used to characterize the shrinkage behaviors for a wide range of commercial and experimental visible-light-cure materials in relation to the composition, boundary condition and cavity geometry.

Image time-correlation, dynamic light scattering, and birefringence for the study of the response of anisometric colloids to external fields.

In this paper, a detailed description of equipment is given, specially designed to characterize the response of non-spherical colloids to external fields. To characterize slow structural changes on a large length scale we developed an image correlation method, fast dynamics on the colloidal-particle level is probed by means of a vertically mounted, small angle dynamic light scattering setup, while the orientational order (induced by the external field) is measured with a birefringence setup with off-normal incidence. The performance of this in situ set of instruments is illustrated by experiments on concentrated dispersions of very long and thin, charged colloidal rods (fd-virus particles) in external electric fields. Here, the frequency of the field is sufficiently low to polarize electrical double layers, leading to additional inter-colloidal interactions which are found to give rise to phase/state transitions and dynamical states (K. Kang and J. K. G. Dhont, Soft Matter 6, 273, 2010).

Constitutive equations for the flow behavior of entangled polymeric systems: application to star polymers.

A semimicroscopic derivation is presented of equations of motion for the density and the flow velocity of concentrated systems of entangled polymers. The essential ingredient is the transient force that results from perturbations of overlapping polymers due to flow. A Smoluchowski equation is derived that includes these transient forces. From this, an equation of motion for the polymer number density is obtained, in which body forces couple the evolution of the polymer density to the local velocity field. Using a semimicroscopic Ansatz for the dynamics of the number of entanglements between overlapping polymers, and for the perturbations of the pair-correlation function due to flow, body forces are calculated for nonuniform systems where the density as well as the shear rate varies with position. Explicit expressions are derived for the shear viscosity and normal forces, as well as for nonlocal contributions to the body force, such as the shear-curvature viscosity. A contribution to the equation of motion for the density is found that describes mass transport due to spatial variation of the shear rate. The two coupled equations of motion for the density and flow velocity predict flow instabilities that will be discussed in more detail in a forthcoming publication.

Diffusion of spheres in isotropic and nematic networks of rods: electrostatic interactions and hydrodynamic screening.

Translational diffusion of a small charged tracer sphere in isotropic and nematic suspensions of long and thin charged rods is investigated as a function of ionic strength and rod concentration. A theory for the diffusive properties of a small sphere is developed, where both (screened) hydrodynamic interactions and charge interactions between the tracer sphere and the rod network are analyzed. Hydrodynamic interactions are formulated in terms of the hydrodynamic screening length. As yet, there are no independent theoretical predictions for the hydrodynamic screening length for rod networks. Experimental tracer-diffusion data are presented for various ionic strengths as a function of the rod concentration, both in the isotropic and nematic states. Orientational order parameters are measured for the same ionic strengths as a function of the rod concentration. The hydrodynamic screening length is determined from these experimental data and scaling relations obtained from the above mentioned theory. For the isotropic networks, a master curve is found for the hydrodynamic screening length as a function of the rod concentration. For the nematic networks the screening length turns out to be a very sensitive function of the orientational order parameter.

Vorticity banding in rodlike virus suspensions.

Vorticity banding under steady shear flow is observed in a suspension of semiflexible colloidal rods (fd virus particles) within a part of the paranematic-nematic biphasic region. Banding occurs uniformly throughout the cell gap within a shear-rate interval (.gamma-, .gamma+) , which depends on the fd concentration. For shear rates below the lower-border shear rate .gamma- only shear elongation of inhomogeneities, which are formed due to paranematic-nematic phase separation, is observed. Within a small region just above the upper-border shear rate .gamma+ , banding occurs heterogeneously. An essential difference in the kinetics of vorticity banding is observed, depending on the morphology of inhomogeneities formed during the initial stages of the paranematic-nematic phase separation. Particle tracking and polarization experiments indicate that the vorticity bands are in a weak rolling flow, superimposed on the applied shear flow. We propose a mechanism for the origin of the banding instability and the transient stability of the banded states. This mechanism is related to the normal stresses generated by inhomogeneities formed due to the underlying paranematic-nematic phase transition.

Diffusion of spheres in isotropic and nematic suspensions of rods.

Diffusion of a small tracer sphere (apoferritin) in isotropic and nematic networks [of fd virus] is discussed. For a tracer sphere that is smaller than the mesh size of the network, screened hydrodynamic interactions between the sphere and the network determine its diffusion coefficient. A theory is developed for such interactions as well as their relation to the long-time self-diffusion coefficient. Fluorescence correlation spectroscopy measurements on mixtures of apoferritin and fd virus are presented. The long-time self-diffusion coefficient of apoferritin is measured as a function of the fd-virus concentration, both in the isotropic and nematic state, in directions parallel and perpendicular to the nematic director. The hydrodynamic screening length of the fd-virus network as a function of fd concentration is obtained by combining these experimental data with the theory. Surprisingly, the screening length increases with increasing concentration in nematic networks. This is due to the increase in the degree of alignment, which apparently leads to a strong increase of the screening length. Hydrodynamic screening is thus strongly diminished by alignment. A self-consistent calculation of the screening length does not work at higher concentrations, probably due to the strong variation of the typical incident flow fields over the contour of a rod.

Nematic-isotropic spinodal decomposition kinetics of rodlike viruses.

We investigate spinodal decomposition kinetics of an initially nematic dispersion of rodlike viruses. Quench experiments are performed from a flow-stabilized homogeneous nematic state at a high shear rate into the two-phase isotropic-nematic coexistence region at a zero shear rate. We present experimental evidence that spinodal decomposition is driven by orientational diffusion, in accordance with a very recent theory.