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Dynamic characterization of hydrophobic and hydrophilic solutes in oleic-acid enhanced transdermal delivery using two-photon fluorescence microscopy

In this letter, we propose an efficient methodology of investigating dynamic properties of sulforhodamine B and rhodamine B hexyl ester molecules transporting across ex-vivo human stratum corneum with and without oleic acid enhancement. Three-dimensional, time-lapse fluorescence images of the stratum corneum can be obtained using two-photon fluorescence microscopy. Furthermore, temporal quantifications of transport enhancements in diffusion parameters can be achieved with the use of Fick's second law. Dynamic characterization of solutes transporting across the stratum corneum is an effective method for understanding transient phenomena in transdermal delivery of probe molecules, leading to improved delivery strategies of molecular species for therapeutic purposes.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Sub-nanometer glass surface dynamics induced by illumination

Illumination is known to induce stress and morphology changes in opaque glasses. Amorphous silicon carbide (a-SiC) has a smaller bandgap than the crystal. Thus, we were able to excite with 532 nm light a 1 μm amorphous surface layer on a SiC crystal while recording time-lapse movies of glass surface dynamics by scanning tunneling microscopy (STM). Photoexcitation of the a-SiC surface layer through the transparent crystal avoids heating the STM tip. Up to 6 × 10{sup 4} s, long movies of surface dynamics with 40 s time resolution and sub-nanometer spatial resolution were obtained. Clusters of ca. 3-5 glass forming units diameter are seen to cooperatively hop between two states at the surface. Photoexcitation with green laser light recruits immobile clusters to hop, rather than increasing the rate at which already mobile clusters hop. No significant laser heating was observed. Thus, we favor an athermal mechanism whereby electronic excitation of a-SiC directly controls glassy surface dynamics. This mechanism is supported by an exciton migration-relaxation-thermal diffusion model. Individual clusters take ∼1 h to populate states differently after the light intensity has changed. We believe the surrounding matrix rearranges slowly when it is stressed by a change in laser intensity, and clusters serve as a diagnostic. Such cluster hopping and matrix rearrangement could underlie the microscopic mechanism of photoinduced aging of opaque glasses.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Live cell imaging combined with high-energy single-ion microbeam

DNA strand breaks can lead to cell carcinogenesis or cell death if not repaired rapidly and efficiently. An online live cell imaging system was established at the high energy microbeam facility at the Institute of Modern Physics to study early and fast cellular response to DNA damage after high linear energy transfer ion radiation. The HT1080 cells expressing XRCC1-RFP were irradiated with single high energy nickel ions, and time-lapse images of the irradiated cells were obtained online. The live cell imaging analysis shows that strand-break repair protein XRCC1 was recruited to the ion hit position within 20 s in the cells and formed bright foci in the cell nucleus. The fast recruitment of XRCC1 at the ion hits reached a maximum at about 200 s post-irradiation and then was followed by a slower release into the nucleoplasm. The measured dual-exponential kinetics of XRCC1 protein are consistent with the proposed consecutive reaction model, and the measurements obtained that the reaction rate constant of the XRCC1 recruitment to DNA strand break is 1.2 × 10{sup −3} s{sup −1} and the reaction rate constant of the XRCC1 release from the break-XRCC1 complex is 1.2 × 10{sup −2} s{sup −1}.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

High-speed atomic force microscopy reveals structural dynamics of α-synuclein monomers and dimers

We report that α-Synuclein (α-syn) is the major component of the intraneuronal inclusions called Lewy bodies, which are the pathological hallmark of Parkinson’s disease. α-Syn is capable of self-assembly into many different species, such as soluble oligomers and fibrils. Even though attempts to resolve the structures of the protein have been made, detailed understanding about the structures and their relationship with the different aggregation steps is lacking, which is of interest to provide insights into the pathogenic mechanism of Parkinson’s disease. Here we report the structural flexibility of α-syn monomers and dimers in an aqueous solution environment as probed by single-molecule time-lapse high-speed AFM. In addition, we present the molecular basis for the structural transitions using discrete molecular dynamics (DMD) simulations. α-Syn monomers assume a globular conformation, which is capable of forming tail-like protrusions over dozens of seconds. Importantly, a globular monomer can adopt fully extended conformations. Dimers, on the other hand, are less dynamic and show a dumbbell conformation that experiences morphological changes over time. DMD simulations revealed that the α-syn monomer consists of several tightly packed small helices. The tail-like protrusions are also helical with a small β-sheet, acting as a “hinge”. Monomers within dimers have a large interfacial interaction area and are stabilized by interactions in the non-amyloid central (NAC) regions. Additionally, the dimer NAC-region of each α-syn monomer forms a β-rich segment. Moreover, NAC-regions are located in the hydrophobic core of the dimer.

59 BASIC BIOLOGICAL SCIENCES↗

Rosette-scan video-rate atomic force microscopy: Trajectory patterning and control design

We introduce an analysis and a systematic design methodology for a novel nonraster scan method based on a rosette pattern and demonstrate its application in video-rate atomic force microscopy. This pattern is traced when the lateral axes of a parallel kinematic scanner are commanded to follow a combination of two sinusoids with identical amplitudes and different frequencies. We create an internal-model-based controller to enhance the tracking performance of this pattern and implement the scheme on a microelectromechanical system scanner. The conclusions reveal high-precision tracking of the rosette pattern in order to acquire time-lapsed atomic force microscope images at the rate of 10 frames/s.

42 ENGINEERING↗

Reconstruction of the interatomic forces from dynamic scanning transmission electron microscopy data

We explore the possibility for reconstruction of the generative physical models describing interactions between atomic units in solids from observational electron microscopy data. In this work, scanning transmission electron microscopy (STEM) is used to observe the dynamic motion of Si atoms at the edge of monolayer graphene under continuous electron beam illumination. The resulting time-lapsed STEM images represent the snapshots of observed chemical states of the system. We use two approaches: potential of mean force calculation using a radial distribution function and a direct fitting of the graphene–Si interatomic pairwise potentials with force matching, to reconstruct the force fields in the materials. These studies lay the foundation for quantitative analysis of materials energetics from STEM data through the sampling of the metastable states in the chemical space of the system.

42 ENGINEERING↗

Formation of an optical pulsed discharge in a supersonic air flow by radiation of a repetitively pulsed CO{sub 2} laser

Results of optimisation of repetitively pulsed CO{sub 2}-laser generation are presented for finding physical conditions of forming stable burning of an optical pulsed discharge (OPD) in a supersonic air flow and for studying the influence of pulse parameters on the energy absorption efficiency of laser radiation in plasma. The optical discharge in a supersonic air flow was formed by radiation of a repetitively pulsed CO{sub 2} laser with mechanical Q-switching excited by a discharge with a convective cooling of the working gas. For the first time the influence of radiation pulse parameters on the ignition conditions and stable burning of the OPD in a supersonic air flow was investigated and the efficiency of laser radiation absorption in plasma was studied. The influence of the air flow velocity on stability of plasma production was investigated. It was shown that stable burning of the OPD in a supersonic flow is realised at a high pulse repetition rate where the interval between radiation pulses is shorter than the time of plasma blowing-off. Study of the instantaneous value of the absorption coefficient shows that after a breakdown in a time lapse of 100 - 150 ns, a quasi-stationary 'absorption phase' is formed with the duration of {approx}1.5 ms, which exists independently of air flow and radiation pulse repetition rate. This phase of strong absorption is, seemingly, related to evolution of the ionisation wave. (laser applications and other topics in quantum electronics)

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Transcriptional burst frequency and burst size are equally modulated across the human genome

Gene expression occurs either as an episodic process, characterized by pulsatile bursts or as a constitutive, Poisson-like accumulation of gene products. It is not clear which mode of gene expression (constitutive versus bursty) predominates across a genome or how transcriptional dynamics are influenced by genomic position and promoter sequence. Here, we use time-lapse fluorescence microscopy, building off of theoretical studies that exploit the time-resolved structure of stochastic fluctuations in gene expression, to develop a three-dimensional method for mapping underlying gene-regulatory mechanisms. Over 8,000 individual human genomic loci were analyzed, and at virtually all loci, episodic bursting as opposed to constitutive expression was found to be the predominant mode of expression. Quantitative analysis of the expression dynamics at these 8,000 loci indicates that both frequency and size of transcriptional bursts vary equally across the human genome independent of promoter sequence. Strikingly, weaker expression loci modulate burst frequency to increase activity, while stronger expression loci modulate burst size to increase activity. Transcriptional activators, such as TNF, generate similar patterns of change in burst frequency and burst size. In summary, transcriptional bursting dominates across the human genome, both burst frequency and burst size vary by chromosomal location, and transcriptional activators alter burst frequency and burst size, depending on the expression level of the locus.

noise biology↗

Superresolution fluorescence mapping of single-nanoparticle catalysts reveals spatiotemporal variations in surface reactivity

For the practical application of nanocatalysts, it is desirable to understand the spatiotemporal fluctuations of nanocatalytic activity at the single-nanoparticle level. Here we use time-lapsed superresolution mapping of single-molecule catalysis events on individual nanoparticles to observe time-varying changes in the spatial distribution of catalysis events on Sb-doped TiO 2 nanorods and Au triangle nanoplates. Compared with the active sites on well-defined surface facets, the defects of the nanoparticle catalysts possess higher intrinsic reactivity but lower stability. Corners and ends are more reactive but also less stable than flat surfaces. Averaged over time, the most stable sites dominate the total apparent activity of single nanocatalysts. However, the active sites with higher intrinsic activity but lower stability show activity at earlier time points before deactivating. Unexpectedly, some active sites are found to recover their activity (“self-healing”) after deactivation, which is probably due to desorption of the adsorbate. Finally, our superresolution measurement of different types of active catalytic sites, over both space and time, leads to a more comprehensive understanding of reactivity patterns and may enable the design of new and more productive heterogeneous catalysts.

36 MATERIALS SCIENCE↗

Antifungal symbiotic peptide NCR044 exhibits unique structure and multifaceted mechanisms of action that confer plant protection

In the indeterminate nodules of a model legume Medicago truncatula, ~700 nodule-specific cysteine-rich (NCR) peptides with conserved cysteine signature are expressed. NCR peptides are highly diverse in sequence, and some of these cationic peptides exhibit antimicrobial activity in vitro and in vivo. However, there is a lack of knowledge regarding their structural architecture, antifungal activity, and modes of action against plant fungal pathogens. Here, the three-dimensional NMR structure of the 36-amino acid NCR044 peptide was solved. This unique structure was largely disordered and highly dynamic with one four-residue α-helix and one three-residue antiparallel β-sheet stabilized by two disulfide bonds. NCR044 peptide also exhibited potent fungicidal activity against multiple plant fungal pathogens, including Botrytis cinerea and three Fusarium spp. It inhibited germination in quiescent spores of B. cinerea. In germlings, it breached the fungal plasma membrane and induced reactive oxygen species. It bound to multiple bioactive phosphoinositides in vitro. Time-lapse confocal and superresolution microscopy revealed strong fungal cell wall binding, penetration of the cell membrane at discrete foci, followed by gradual loss of turgor, subsequent accumulation in the cytoplasm, and elevated levels in nucleoli of germlings. Spray-applied NCR044 significantly reduced gray mold disease symptoms caused by the fungal pathogen B. cinerea in tomato and tobacco plants, and postharvest products. Our work illustrates the antifungal activity of a structurally unique NCR peptide against plant fungal pathogens and paves the way for future development of this class of peptides as a spray-on fungistat/fungicide.

59 BASIC BIOLOGICAL SCIENCES↗

THE 1998 NOVEMBER 14 OCCULTATION OF GSC 0622-00345 BY SATURN. II. STRATOSPHERIC THERMAL PROFILE, POWER SPECTRUM, AND GRAVITY WAVES

On 1998 November 14, Saturn and its rings occulted the star GSC 0622-00345. The occultation latitude was 55.{sup 0}5 S. This paper analyzes the 2.3 {mu}m light curve derived by Harrington and French. A fixed-baseline isothermal fit to the light curve has a temperature of 140 {+-} 3 K, assuming a mean molecular mass of 2.35 AMU. The thermal profile obtained by numerical inversion is valid between 1 and 60 {mu}bar. The vertical temperature gradient is > 0.2 K km{sup -1} more stable than the adiabatic lapse rate, but it still shows the alternating-rounded-spiked features seen in many temperature gradient profiles from other atmospheric occultations and usually attributed to breaking gravity (buoyancy) waves. We conduct a wavelet analysis of the thermal profile, and show that, even with our low level of noise, scintillation due to turbulence in Earth's atmosphere can produce large temperature swings in light-curve inversions. Spurious periodic features in the 'reliable' region of a wavelet amplitude spectrum can exceed 0.3 K in our data. We also show that gravity-wave model fits to noisy isothermal light curves can lead to convincing wave 'detections'. We provide new significance tests for localized wavelet amplitudes, wave model fits, and global power spectra of inverted occultation light curves by assessing the effects of pre- and post-occultation noise on these parameters. Based on these tests, we detect several significant ridges and isolated peaks in wavelet amplitude, to which we fit a gravity wave model. We also strongly detect the global power spectrum of thermal fluctuations in Saturn's atmosphere, which resembles the 'universal' (modified Desaubies) curve associated with saturated spectra of propagating gravity waves on Earth and Jupiter.

79 ASTRONOMY AND ASTROPHYSICS↗

X-RAY FADING AND EXPANSION IN THE “MINIATURE SUPERNOVA REMNANT” OF GK PERSEI

We report on a second epoch of Chandra X-ray imaging spectroscopy of the spatially resolved old nova remnant GK Persei. An ACIS-S3 observation of 97.4 ks was conducted in 2013 November after a lapse of 13.8 yr from the last visit in 2000. The X-ray emitting nebula appeared more faint and patchy compared with the first epoch. The flux decline was particularly evident in fainter regions and the mean decline was 30%–40% in the 0.5–1.2 keV energy band. A typical expansion of the brightest part of the remnant was 1.″9, which corresponds to an expansion rate of 0.″14 yr{sup −1}. The soft X-ray spectra extracted from both the 2000 and 2013 data can be explained by a non-equilibrium ionization collisional plasma model convolved with interstellar absorption, though do not allow us to constrain the origin of the flux evolution. The plasma temperature has not significantly evolved since the 2000 epoch and we conclude that the fading of the X-ray emission is due largely to expansion. This implies that recent expansion has been into a lower density medium, a scenario that is qualitatively consistent with the structure of the circumstellar environment photographed soon after the initial explosion more than a century ago. Fainter areas are fading more quickly than brighter areas, indicating that they are fainter because of a lower ambient medium density and consequently more rapid expansion.

79 ASTRONOMY AND ASTROPHYSICS↗

Essential building blocks of dark energy

We propose a minimal description of single field dark energy/modified gravity within the effective field theory formalism for cosmological perturbations, which encompasses most existing models. We start from a generic Lagrangian given as an arbitrary function of the lapse and of the extrinsic and intrinsic curvature tensors of the time hypersurfaces in unitary gauge, i.e. choosing as time slicing the uniform scalar field hypersurfaces. Focusing on linear perturbations, we identify seven Lagrangian operators that lead to equations of motion containing at most two (space or time) derivatives, the background evolution being determined by the time-dependent coefficients of only three of these operators. We then establish a dictionary that translates any existing or future model whose Lagrangian can be written in the above form into our parametrized framework. As an illustration, we study Horndeski's — or generalized Galileon — theories and show that they can be described, up to linear order, by only six of the seven operators mentioned above. This implies, remarkably, that the dynamics of linear perturbations can be more general than that of Horndeski while remaining second order. Finally, in order to make the link with observations, we provide the entire set of linear perturbation equations in Newtonian gauge, the effective Newton constant in the quasi-static approximation and the ratio of the two gravitational potentials, in terms of the time-dependent coefficients of our Lagrangian.

79 ASTRONOMY AND ASTROPHYSICS↗

Translation invariant time-dependent solutions to massive gravity II

This paper is a sequel to JCAP 12 (2013) 004 and is also devoted to translation-invariant solutions of ghost-free massive gravity in its moving frame formulation. Here we consider a mass term which is linear in the vielbein (corresponding to a β{sub 3} term in the 4D metric formulation) in addition to the cosmological constant. We determine explicitly the constraints, and from the initial value formulation show that the time-dependent solutions can have singularities at a finite time. Although the constraints give, as in the β{sub 1} case, the correct number of degrees of freedom for a massive spin two field, we show that the lapse function can change sign at a finite time causing a singular time evolution. This is very different to the β{sub 1} case where time evolution is always well defined. We conclude that the β{sub 3} mass term can be pathological and should be treated with care.

79 ASTRONOMY AND ASTROPHYSICS↗

Translation invariant time-dependent massive gravity: Hamiltonian analysis

The canonical structure of the massive gravity in the first order moving frame formalism is studied. We work in the simplified context of translation invariant fields, with mass terms given by general non-derivative interactions, invariant under the diagonal Lorentz group, depending on the moving frame as well as a fixed reference frame. We prove that the only mass terms which give 5 propagating degrees of freedom are the dRGT mass terms, namely those which are linear in the lapse. We also complete the Hamiltonian analysis with the dynamical evolution of the system.

79 ASTRONOMY AND ASTROPHYSICS↗

'Constraint consistency' at all orders in cosmological perturbation theory

We study the equivalence of two—order-by-order Einstein's equation and Reduced action—approaches to cosmological perturbation theory at all orders for different models of inflation. We point out a crucial consistency check which we refer to as 'Constraint consistency' condition that needs to be satisfied in order for the two approaches to lead to identical single variable equation of motion. The method we propose here is quick and efficient to check the consistency for any model including modified gravity models. Our analysis points out an important feature which is crucial for inflationary model building i.e., all 'constraint' inconsistent models have higher order Ostrogradsky's instabilities but the reverse is not true. In other words, one can have models with constraint Lapse function and Shift vector, though it may have Ostrogradsky's instabilities. We also obtain single variable equation for non-canonical scalar field in the limit of power-law inflation for the second-order perturbed variables.

79 ASTRONOMY AND ASTROPHYSICS↗

Effective description of higher-order scalar-tensor theories

Most existing theories of dark energy and/or modified gravity, involving a scalar degree of freedom, can be conveniently described within the framework of the Effective Theory of Dark Energy, based on the unitary gauge where the scalar field is uniform. We extend this effective approach by allowing the Lagrangian in unitary gauge to depend on the time derivative of the lapse function. Although this dependence generically signals the presence of an extra scalar degree of freedom, theories that contain only one propagating scalar degree of freedom, in addition to the usual tensor modes, can be constructed by requiring the initial Lagrangian to be degenerate. Starting from a general quadratic action, we derive the dispersion relations for the linear perturbations around Minkowski and a cosmological background. Our analysis directly applies to the recently introduced Degenerate Higher-Order Scalar-Tensor (DHOST) theories. For these theories, we find that one cannot recover a Poisson-like equation in the static linear regime except for the subclass that includes the Horndeski and so-called 'beyond Horndeski' theories. We also discuss Lorentz-breaking models inspired by Horava gravity.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

A class of minimally modified gravity theories

We investigate the Hamiltonian structure of a class of gravitational theories whose actions are linear in the lapse function. We derive the necessary and sufficient condition for a theory in this class to have two or less local physical degrees of freedom. As an application we then find several concrete examples of modified gravity theories in which the total number of local physical degrees of freedom in the gravity sector is two.

79 ASTRONOMY AND ASTROPHYSICS↗