Properties of twisted topological defects in 2D nematic liquid crystals.

Topological defects are one of the most conspicuous features of liquid crystals. In two dimensional nematics, they have been shown to behave effectively as particles with both charge and orientation, which dictate their interactions. Here, we study "twisted" defects that have a radially dependent orientation. We find that twist can be partially relaxed through the creation and annihilation of defect pairs. By solving the equations for defect motion and calculating the forces on defects, we identify four distinct elements that govern the relative relaxational motion of interacting topological defects, namely attraction, repulsion, co-rotation and co-translation. The interaction of these effects can lead to intricate defect trajectories, which can be controlled by setting relevant timescales.

Versatile optical laser system for experiments at the European X-ray free-electron laser facility.

We present the main features of the final prototype of a pulsed optical laser, developed for pump-probe and other experiments in conjunction with the femtosecond x-ray beams at the European X-ray free-electron laser facility. Adapted to the temporal x-ray emission pattern of the facility, the laser provides 10 Hz bursts of up to 600 µs duration with intra-burst pulse frequencies as high as 4.5 MHz. In this mode, we have generated pulses as short as 12 fs at 350 W average power during the burst and with beam qualities close to the diffraction limit. This is, to the best of our knowledge, the highest power to date of a few-cycle laser operating at a center wavelength of 800 nm. Important for experimental flexibility, the laser can be configured in various unique ways, enabling, e.g., energy scaling to >3 mJ per pulse through a frequency change down to 100 kHz and the generation of nearly transform limited pulses between 12 fs and 300 fs. In addition to the 800 nm femtosecond beam line, a synchronized long pulse (0.8 ps or 400 ps) 1030 nm beam can be utilized, offering up to 4 kW burst average power, i.e. up to 40 mJ per pulse at 100 kHz. Efficient nonlinear wavelength conversion and tuning through intrinsic and external means further enhance the capabilities of the laser.

Generation of stable overlaps between antiparallel filaments.

During cell division, sister chromatids are segregated by the mitotic spindle, a bipolar assembly of interdigitating antiparallel polar filaments called microtubules. Establishing a stable overlap region is essential for maintenance of bipolarity, but the underlying mechanisms are poorly understood. Using a particle-based stochastic model, we find that the interplay of motors and passive cross-linkers can robustly generate partial overlaps between antiparallel filaments. In this situation, motors reduce the overlap in a length-dependent manner, whereas passive cross-linkers increase it independently of the length. In addition to maintaining structural integrity, passive cross-linkers can thus also have a dynamic role for overlap size regulation.

High power burst-mode optical parametric amplifier with arbitrary pulse selection.

We present results from a unique burst-mode femtosecond non-collinear optical parametric amplifier (NOPA) under development for the optical - x-ray pump-probe experiments at the European X-Ray Free-Electron Laser Facility. The NOPA operates at a burst rate of 10 Hz, a duty cycle of 2.5% and an intra-burst repetition rate of up to 4.5 MHz, producing high fidelity 15 fs pulses at a center wavelength of 810 nm. Using dispersive amplification filtering of the super-continuum seed pulses allows for selectable pulse duration up to 75 fs, combined with a tuning range in excess of 100 nm whilst remaining nearly transform limited. At an intra-burst rate of 188 kHz the single pulse energy from two sequential NOPA stages reached 180 µJ, corresponding to an average power of 34W during the burst. Acousto- and electro-optic switching techniques enable the generation of transient free bursts of required length and the selection of arbitrary pulse sequences inside the burst.

The actin cortex as an active wetting layer.

Using active gel theory we study theoretically the properties of the cortical actin layer of animal cells. The cortical layer is described as a non-equilibrium wetting film on the cell membrane. The actin density is approximately constant in the layer and jumps to zero at its edge. The layer thickness is determined by the ratio of the polymerization velocity and the depolymerization rate of actin.

Treadmilling and length distributions of active polar filaments.

The cytoskeleton is a network of filamentous proteins, notably, actin filaments and microtubules. These filaments are active as their assembly is driven by the hydrolysis of nucleotides bound to the constituting protomers. In addition, the assembly kinetics differs at the two respective ends, making them active polar filaments. Experimental evidence suggests, that, in vivo, actin filaments and microtubules can grow at one and shrink at the other end at the same rate, a state that is known as treadmilling. In this work, we use a generic discrete two-state model for active polar filaments to analyze the conditions leading to treadmilling. We find that a single filament can self-organize into the treadmilling state for a broad range of monomer concentrations. In this regime the corresponding length distribution has a pronounced maximum at a finite value. We then extend our description to consider specifically the dynamics of actin filaments. We show that actin treadmilling should be observable in vitro in the presence of appropriate depolymerization promoting factors.

Cell motility resulting from spontaneous polymerization waves.

The crawling of cells on a substrate is in many cases driven by the actin cytoskeleton. How actin filaments and associated proteins are organized to generate directed motion is still poorly understood. Recent experimental observations suggest that spontaneous cytoskeletal waves might orchestrate the actin-filament network to produce directed motion. We investigate this possibility by studying a mean-field description of treadmilling filaments interacting with nucleating proteins, a system that is known to self-organize into waves. Confining the system by a boundary that shares essential features of membranes, we find that spontaneous waves can generate directional motion. We also find that it can produce lateral waves along the confining membrane as are observed in spreading cells.

Self-organization in systems of treadmilling filaments.

The cytoskeleton is an important substructure of living cells, playing essential roles in cell division, cell locomotion, and the internal organization of subcellular components. Physically, the cytoskeleton is an active polar gel, that is, a system of polar filamentous polymers, which is intrinsically out of thermodynamic equilibrium. Active processes are notably involved in filament growth and can lead to net filament assembly at one end and disassembly at the other, a phenomenon called treadmilling. Here, we develop a framework for describing collective effects in systems of treadmilling filaments in the presence of agents regulating filament assembly. We find that such systems can self-organize into asters and moving filament blobs. We discuss possible implications of our findings for subcellular processes.

Evaluation of potential carryover effects associated with limit feeding of gravid Holstein heifers.

Ninety-six Holstein heifers (400±6kg, 15.2±0.1 mo), including 9 with ruminal cannulas, were offered 1 of 3 diets for 180±8 d in a randomized replicated pen design. Dietary treatments included a control diet (C100) and 2 independent limit-fed (LF) diets. The LF diets included one offered at 85% of C100 intake (L85) without an ionophore, and a second containing an ionophore (325 mg/head per day of lasalocid) that was offered at 80% of C100 intake (L80+I). Heifers were evaluated for growth, rumen digesta volume, nutrient excretion, and subsequent lactation performance. Limit-fed heifers consumed less dry matter and neutral detergent fiber, and had greater respective average daily gains (0.96 or 0.89 vs. 0.81 kg/d), and lower feed:gain ratios (9.1 or 9.3 vs. 13.0 kg/kg) compared with heifers offered the C100 diet. No differences in rumen pH, NH(3)-N, or volatile fatty acid concentrations were observed between C100 and LF heifers. Rumen digesta volume, density, and weight were unaffected by LF, and feeding L85 or L80+I did not result in carryover effects for rumen digesta volume when these heifers were offered a common high-fiber diet immediately after the 180-d growth trial. At parturition, no differences were observed for dystocia index, calf body weight, or 7-d postpartum body weight between cows offered LF or C100 diets as heifers. Lactation body weight, dry matter intake, and feed efficiency of cows did not differ between treatments at 45 or 90 d in milk. Milk yield and milk components also were not different between cows that were offered C100 or LF diets as gravid heifers. At 45 d in milk, rumen digesta volume was greater (99.1 vs. 66.1L) for cows offered L85 compared with cows offered L80+I as gravid heifers, but this effect was not observed at 90 d in milk. Limit feeding of gravid Holstein heifers for 180 d did not result in any carryover effects during their first lactation for rumen digesta volume, dry matter intake, or milk yield.

Uncorrelated changes of subunit stability can generate length-dependent disassembly of treadmilling filaments.

Unregulated polymer assembly typically leads to exponential length distributions. In living cells, though, the length distribution of cytoskeletal filaments often shows a sharp maximum at a finite value. We discuss a simple mechanism for length regulation that is based on generating an effectively length-dependent disassembly rate by locally acting cytoskeletal proteins. We analyze steady-state distributions by stochastic simulations and develop a method to analytically characterize the typical filament length. We find three different types of behavior: (1) unbounded filament growth, (2) exponential distributions and (3) unimodal distributions, which can be very sharp with a standard deviation much smaller than the mean filament length. We discuss possible implications of our results for subcellular processes.

A constitutively active mutant PTH-PTHrP receptor in Jansen-type metaphyseal chondrodysplasia.

A single heterozygous nucleotide exchange in exon M2 of the gene encoding the parathyroid hormone-parathyroid hormone-related peptide (PTH-PTHrP) receptor was identified in a patient with Jansen-type metaphyseal chondrodysplasia, which changes a strictly conserved histidine residue at position 223 in the receptor's first intracellular loop to arginine. Constitutive, ligand-independent adenosine 3',5'-monophosphate accumulation was observed in COS-7 cells expressing the mutant PTH-PTHrP receptor but not in cells expressing the wild-type receptor. This finding explains the severe ligand-independent hypercalcemia and hypophosphatemia, and most likely the abnormal formation of endochondral bone, in this rare form of short-limbed dwarfism.

Quantitative determination of airborne respirable non-fibrous alpha-silicon carbide by x-ray powder diffractometry.

OBJECTIVES: The purpose of the present investigation was to establish a method for the determination of airborne respirable non-fibrous silicon carbide (SiC). The main application is within the industrial production of SiC. METHODS: Due to the complex airborne aerosol mixture of crystalline compounds in the SiC industry, X-ray powder diffractometry was selected as the most appropriate method. Without any international standard material for the respirable fraction of non-fibrous SiC, pure and suitable products from three SiC plants in Norway were selected. These products have a median particle diameter in the range 4.4-5.1 mum. The method is based on thin sample technique, with the dust deposited on a polycarbonate filter. Absorption correction is done by standard procedures with the use of a silver filter, situated below the polycarbonate filter. RESULTS: The diffraction line used for quantitative determination was selected carefully. This was done to avoid interferences from quartz, cristobalite, and graphite, which all are airborne components present in the atmosphere during the industrial process. The instrumental limit of detection for the method is 12 microg. CONCLUSIONS: This method has been used to determine airborne non-fibrous SiC in a comprehensive ongoing project in the Norwegian SiC industry for further epidemiological studies. The method is fully applicable for compliance work.

Generation and homogeneity of aerosols in a human whole-body inhalation chamber.

INTRODUCTION: A 16 m(3) whole-body exposure chamber for human exposure to aerosols is described. Several modifications of the aerosol generation and distribution system were needed to ensure a stable aerosol concentration in the chamber, especially when a cyclone pre-classifier was used. RESULTS: After these modifications, stable aerosol concentrations of aluminium oxide with a volume median diameter of 5.7 microm, and approximately 3 microm when the cyclone was used, could be achieved after 1 h of aerosol generation. Aerosol concentrations of 1-8 mg m(-3) generated without the cyclone could be maintained for at least 2 h after the aerosol level had stabilized. The temporal variability [coefficient of variation (CV)] of the aerosol concentration was 4-6%, while concentrations <1 mg m(-3) showed greater relative variability. The spatial variability at 3.8 mg m(-3) without a volunteer in the chamber was 4.8%. With a volunteer in the chamber who performed 30 min of ergometric cycling during 2 h of aerosol exposure, the exposure estimated by personal sampling was 15-17% lower than monitored with an optical particle counter. The variability of personally measured exposure was higher than of stationary measurements showing CVs of 10-19%. CONCLUSIONS: These results show that controlled exposure of human volunteers to a range of concentrations can be achieved with good accuracy in this inhalation chamber. The results compare favourably with other chambers described in the literature. Personal sampling showed lower aerosol concentrations than estimated in an empty chamber and the variability was significantly higher than measured stationary.

Anomalous percolation features in molecular evolution.

Self-replication underlies every species of living beings and simple physical intuition dictates that some sort of autocatalysis invariably constitutes a necessary ingredient for the emergence of molecular life. This led Worst et al. [E. G. Worst, P. Zimmer, E. Wollrab, K. Kruse, and A. Ott, New J. Phys. 18, 103003 (2016)NJOPFM1367-263010.1088/1367-2630/18/10/103003] to study a model of molecular evolution of self-replicating molecules where spontaneous ligation and simple autocatalysis are in competition for their building blocks. We revisit this model, where irreversible aggregation leads to a transition from a regime of small molecules to macromolecules, and find an array of anomalous percolation features, some of them predicted for very specific percolation processes [R. M. D'Souza and J. Nagler, Nat. Phys. 11, 531 (2015)1745-247310.1038/nphys3378].

Active bundles of polar and bipolar filaments.

Bundles of actin filaments and molecular motors of the myosin family are a common subcellular organizational motif. Typically, such bundles are under contractile stress resulting from interactions between the filaments and the motors. This holds in particular for contractile rings that appear in the late stages of cell division in animal cells and that cleave the mother into two daughter cells. It was recently shown that myosin organizes into regularly spaced clusters along rings in mammalian cells, whereas myosin clusters in fission yeast travel along the perimeter of actomyosin rings [Wollrab et al., Nat. Commun. 7, 11860 (2016)2041-172310.1038/ncomms11860]. A mechanism based on the association of the structurally polar actin filaments into bipolar structures was shown to provide a common explanation for both observations. Here, we analyze the dynamics of this mechanism in detail. We find a rich phase diagram depending on the actomyosin interaction strength and the stability of the bipolar structures. The system can notably organize into traveling waves. Furthermore, we identify the nature of the bifurcations connecting the various patterns as parameters are changed. Finally, we report experimental patterns observed in cytokinetic rings in fission yeast and link them to solutions of our dynamic equations. Our analysis highlights the possible role played by local polarity sorting of actin filaments for the dynamics and functionality of actomyosin networks.

Spontaneous buckling of contractile poroelastic actomyosin sheets.

Shape transitions in developing organisms can be driven by active stresses, notably, active contractility generated by myosin motors. The mechanisms generating tissue folding are typically studied in epithelia. There, the interaction between cells is also coupled to an elastic substrate, presenting a major difficulty for studying contraction induced folding. Here we study the contraction and buckling of active, initially homogeneous, thin elastic actomyosin networks isolated from bounding surfaces. The network behaves as a poroelastic material, where a flow of fluid is generated during contraction. Contraction starts at the system boundaries, proceeds into the bulk, and eventually leads to spontaneous buckling of the sheet at the periphery. The buckling instability resulted from system self-organization and from the spontaneous emergence of density gradients driven by the active contractility. The buckling wavelength increases linearly with sheet thickness. Our system offers a well-controlled way to study mechanically induced, spontaneous shape transitions in active matter.

Molecular basis for defective secretion of the Z variant of human alpha-1-proteinase inhibitor: secretion of variants having altered potential for salt bridge formation between amino acids 290 and 342.

Human alpha-1-proteinase inhibitor (A1PI) deficiency, associated with the Z-variant A1PI (A1PI/Z) gene, results from defective secretion of the inhibitor from the liver. The A1PI/Z gene exhibits two point mutations which specify amino acid substitutions, Val-213 to Ala and Glu-342 to Lys. The functional importance of these substitutions in A1PI deficiency was investigated by studying the secretion of A1PI synthesized in COS cells transfected with A1PI genes altered by site-directed mutagenesis. This model system correctly duplicates the secretion defect seen in individuals homozygous for the A1PI/Z allele and shows that the substitution of Lys for Glu-342 alone causes defective secretion of A1PI. The substitution of Lys for Glu-342 eliminates the possibility for a salt bridge between residues 342 and 290, which may decrease the conformational stability of the molecule and thus account for the secretion defect. However, when we removed the potential to form a salt bridge from the wild-type inhibitor by changing Lys-290 to Glu (A1PI/SB-290Glu), secretion was not reduced to the 19% of normal level seen for A1PI/Z-342Lys; in fact, 75% of normal secretion was observed. When the potential for salt bridge formation was returned to A1PI/Z-342Lys by changing Lys-290 to Glu, only 46% of normal secretion was seen. These data indicate that the amino acid substitution at position 342, rather than the potential to form the 290-342 salt bridge, is the critical alteration leading to the defect in A1PI secretion.

Morphogen transport in epithelia.

We present a general theoretical framework to discuss mechanisms of morphogen transport and gradient formation in a cell layer. Trafficking events on the cellular scale lead to transport on larger scales. We discuss in particular the case of transcytosis where morphogens undergo repeated rounds of internalization into cells and recycling. Based on a description on the cellular scale, we derive effective nonlinear transport equations in one and two dimensions which are valid on larger scales. We derive analytic expressions for the concentration dependence of the effective diffusion coefficient and the effective degradation rate. We discuss the effects of a directional bias on morphogen transport and those of the coupling of the morphogen and receptor kinetics. Furthermore, we discuss general properties of cellular transport processes such as the robustness of gradients and relate our results to recent experiments on the morphogen Decapentaplegic (Dpp) that acts in the wing disk of the fruit fly Drosophila.

Selective protein degradation in the yeast exocytic pathway.

Protein degradation in the exocytic pathway was studied in Saccharomyces cerevisiae using human alpha-1-protease inhibitor (A1Pi) as a reporter molecule. Yeast cells transformed with A1Pi cDNA genes synthesized A1Pi that entered the secretion pathway and accumulated in the endoplasmic reticulum (ER). Cells expressing A1PiM (wild-type) accumulated about 10-fold more A1Pi than cells expressing A1PiZ (secretion defective variant). Analyses of A1Pi mRNA indicated that the low level of A1PiZ relative to A1PiM was not the result of differential gene transcription. Pulse-chase A1Pi radiolabeling showed that A1PiM and A1PiZ were degraded at different rates and suggested a rapid specific turnover of newly synthesized A1PiZ in the ER. Accumulated A1Pi was degraded at comparable rates in both wild-type cells and cells deficient in vacuolar protease activity, indicating that degradation of A1Pi did not occur in the vacuole. Studies to investigate the intracellular location of the degradative process, using temperature-sensitive secretion defective yeast strains, suggested the possibility that degradation occurs not only in the ER but at a second site accessed by vesicle transport. Together, these results demonstrate that a selective protein degradation process operates early in the yeast cell exocytic pathway.