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At least 55 records · Page 3

The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi

Carbohydrate active enzymes (CAZymes) are vital for the lignocellulose-based biorefinery. The development of hypersecreting fungal protein production hosts is therefore a major aim for both academia and industry. However, despite advances in our understanding of their regulation, the number of promising candidate genes for targeted strain engineering remains limited. Here, we resequenced the genome of the classical hypersecreting Neurospora crassa mutant exo-1 and identified the causative point of mutation to reside in the F-box protein–encoding gene, NCU09899. The corresponding deletion strain displayed amylase and invertase activities exceeding those of the carbon catabolite derepressed strain ?cre-1, while glucose repression was still mostly functional in ?exo-1. Surprisingly, RNA sequencing revealed that while plant cell wall degradation genes are broadly misexpressed in ?exo-1, only a small fraction of CAZyme genes and sugar transporters are up-regulated, indicating that EXO-1 affects specific regulatory factors. Aiming to elucidate the underlying mechanism of enzyme hypersecretion, we found the high secretion of amylases and invertase in ?exo-1 to be completely dependent on the transcriptional regulator COL-26. Furthermore, misregulation of COL-26, CRE-1, and cellular carbon and nitrogen metabolism was confirmed by proteomics. Finally, we successfully transferred the hypersecretion trait of the exo-1 disruption by reverse engineering into the industrially deployed fungus Myceliophthora thermophila using CRISPR-Cas9. Our identification of an important F-box protein demonstrates the strength of classical mutants combined with next-generation sequencing to uncover unanticipated candidates for engineering. These data contribute to a more complete understanding of CAZyme regulation and will facilitate targeted engineering of hypersecretion in further organisms of interest.

Gabriel, Raphael↗

Exfoliated YBCO filaments for second-generation superconducting cable

The second-generation high temperature superconductor (2G HTS) wire is the most enticing conductor for high-field magnets such as accelerator dipoles and compact fusion devices. The key element of the wire is a thin Y 1 Ba 2 Cu 3 O 7 (YBCO) layer deposited on a flexible metal substrate. The substrate, which becomes incorporated in the 2G conductor, reduces the electrical and mechanical performance of the wire. This is a process that exfoliates the YBCO layer from the substrate while retaining the critical current density of the superconductor. Ten-centimeter long coupons of exfoliated YBCO layers were manufactured, and detailed structural, electrical, and mechanical characterization were reported here. After exfoliation, the YBCO layer was supported by a 75 μ m thick stainless steel foil, which makes for a compact, mechanically stronger, and inexpensive conductor. The critical current density of the filaments was measured at both 77 K and 4.2 K. The exfoliated YBCO retained 90% of the original critical current. Similarly, tests in an external magnetic field at 4.2 K confirmed that the pinning strength of the YBCO layer was also retained following exfoliation.

2G conductors↗

Observations from Hinode and SDO of a Twisting and Writhing Start to a Solar-filament-eruption Cascade

Active region eruption of 1 June 2011. Ejective eruption. GOES class C4.1 flare. SDO/AIA, various filters (94, 131, 171, 193, 211, 304, 335 Ang.) High time cadence (24 s) and high spatial resolution (0 .6 pixels). SDO/HMI line-of-sight magnetograms. Hinode observed the onset, and the later decay phase. There are two filament eruptions (filament 1 and filament 2). Filament 1 has slow rise with steps, as in several previous cases. GOES "episodes" play role of "microflares" in other events; that is, filament jumps <=> intensity peaks. Episode 1 brightening: Accompanied by filament 1 s initial motions. (Rest of talk.) Filament 1 becomes unstable, and.. Episode 2 brightening: Flare ribbons following filament 1 s fast liftoff. This destabilizes neighboring filament 2, and... Episode 3 brightening: Flare ribbons of whole system following filament 2 s eruption.Something leads to reconnection; not totally clear what. Reconnection -> twisted flux rope in approx.20 min; episode 1 microflare (flare ribbons; TC) and filament jump. Twist -> writhe, via kink instability; filament-trajectory plateau, approx. 20 min. Writhe -> jump and eruption of filament 1, via instability; episode 2 microflare (flare ribbons; TC). (E.g., Williams et al.) First eruption -> second filament eruption (episode 3 flare ribbons; TC). (E.g., Sterling, Moore; Liu et al.; Torok et al.; Schrijver & Title.). Estimate amount of free energy in newly-twisted field (cf. Moore 1988): where we have taken L and r = 50, 3 arcsec. Energy of the total system is likely 1030 ergs or more. So "no" is answer to question. Additional energy comes from remainder of sheared large loop, shear (free energy) of second filament, etc. (Normally assumed situation.) Some history of twist-induced instability in filament eruptions: e.g., Sakurai, Torok & Kliem, Fan & Gibson, Gilbert et al., van Driel-Gesztelyi et al. Criterion : Kink instability for line-tied tube (Hood & Priest): 2.5pi; for Titov & Demoulin loop (Torok et al): approx.3.5pi We observe here: approx.1.5 turns (3.0pi) over ~50. => consistent with kink instability acting. (Cf. Srivastava et al. (2010): Small flare seen in TRACE and Hinode: approx.6.0pi)

Sterling, Alphonse C.↗

Planck intermediate results: XXXIII. Signature of the magnetic field geometry of interstellar filaments in dust polarization maps

Planck observations at 353 GHz provide the first fully sampled maps of the polarized dust emission towards interstellar filaments and their backgrounds (i.e., the emission observed in the surroundings of the filaments). The data allow us to determine the intrinsic polarization properties of the filaments and therefore to provide insight into the structure of their magnetic field (B). In this paper, we present the polarization maps of three nearby (several parsecs long) star-forming filaments of moderate column density (N H about 10 22 cm -2 ): Musca, B211, and L1506. These three filaments are detected above the background in dust total and polarized emission. We use the spatial information to separate Stokes I, Q, and U of the filaments from those of their backgrounds, an essential step in measuring the intrinsic polarization fraction (p) and angle (ψ) of each emission component. We find that the polarization angles in the three filaments (ψ fil ) are coherent along their lengths and not the same as in their backgrounds (ψ bg ). The differences between ψ fil and ψ bg are 12° and 54° for Musca and L1506, respectively, and only 6° in the case of B211. These differences forMusca and L1506 are larger than the dispersions of ψ, both along the filaments and in their backgrounds. The observed changes of ψ are direct evidence of variations of the orientation of the plane of the sky (POS) projection of the magnetic field. As in previous studies, we find a decrease of several per cent in p with N H from the backgrounds to the crest of the filaments. We show that the bulk of the drop in p within the filaments cannot be explained by random fluctuations of the orientation of the magnetic field because they are too small (σ ψ < 10°). We recognize the degeneracy between the dust alignment efficiency (by, e.g., radiative torques) and the structure of the B-field in causing variations in p, but we argue that the decrease in p from the backgrounds to the filaments results in part from depolarization associated with the 3D structure of the B-field: both its orientation in the POS and with respect to the POS. We do not resolve the inner structure of the filaments, but at the smallest scales accessible with Planck (~0.2 pc), the observed changes of ψ and p hold information on the magnetic field structure within filaments. Finally, they show that both the mean field and its fluctuations in the filaments are different from those of their backgrounds, which points to a coupling between the matter and the B-field in the filament formation process.

79 ASTRONOMY AND ASTROPHYSICS↗

Dependence of thick filament structure in relaxed mammalian skeletal muscle on temperature and interfilament spacing

Contraction of skeletal muscle is regulated by structural changes in both actin-containing thin filaments and myosin-containing thick filaments, but myosin-based regulation is unlikely to be preserved after thick filament isolation, and its structural basis remains poorly characterized. Here, we describe the periodic features of the thick filament structure in situ by high-resolution small-angle x-ray diffraction and interference. We used both relaxed demembranated fibers and resting intact muscle preparations to assess whether thick filament regulation is preserved in demembranated fibers, which have been widely used for previous studies. We show that the thick filaments in both preparations exhibit two closely spaced axial periodicities, 43.1 nm and 45.5 nm, at near-physiological temperature. The shorter periodicity matches that of the myosin helix, and x-ray interference between the two arrays of myosin in the bipolar filament shows that all zones of the filament follow this periodicity. The 45.5-nm repeat has no helical component and originates from myosin layers closer to the filament midpoint associated with the titin super-repeat in that region. Cooling relaxed or resting muscle, which partially mimics the effects of calcium activation on thick filament structure, disrupts the helical order of the myosin motors, and they move out from the filament backbone. Compression of the filament lattice of demembranated fibers by 5% Dextran, which restores interfilament spacing to that in intact muscle, stabilizes the higher-temperature structure. The axial periodicity of the filament backbone increases on cooling, but in lattice-compressed fibers the periodicity of the myosin heads does not follow the extension of the backbone. Thick filament structure in lattice-compressed demembranated fibers at near-physiological temperature is similar to that in intact resting muscle, suggesting that the native structure of the thick filament is largely preserved after demembranation in these conditions, although not in the conditions used for most previous studies with this preparation.

59 BASIC BIOLOGICAL SCIENCES↗

Langmuir turbulence and filament frontogenesis in the oceanic surface boundary layer

Submesoscale currents, small-scale turbulence and surface gravity waves co-exist in the upper ocean and interact in complex ways. To expose the couplings, the frontogenetic life cycle of an idealized cold dense submesoscale filament interacting with upper ocean Langmuir turbulence is investigated in large-eddy simulations (LESs) based on the incompressible wave-averaged equations. The simulations utilize large domains and fine meshes with $6.4\times 10^{9}$ grid points. Case studies are made with surface winds or surface cooling with waves oriented in across-filament (perpendicular) or down-filament (parallel) directions relative to the two-dimensional filament axis. The currents $u$ , $v$ and $w$ are aligned with the across-filament, down-filament and vertical directions, respectively. Frontogenesis is induced by across-filament Lagrangian secondary circulations in the boundary layer, and it is shown to be strongly impacted by surface waves, in particular the propagation direction relative to the filament axis. In a horizontally heterogeneous boundary layer, surface waves induce both mean and fluctuating Stokes-drift vortex forces that modify a linear, hydrostatic turbulent thermal wind (TTW) approximation for momentum. Down-filament winds and waves are found to be especially impactful, they significantly reduce the peak level of frontogenesis by fragmenting the filament into primary and secondary down-welling sites in a broad frontal zone over a width ${\sim}500~\text{m}$ . At peak frontogenesis, opposing down-filament jets $\langle v\rangle$ overlie each other resulting in a vigorous vertical shear layer $\unicode[STIX]{x2202}_{z}\langle v\rangle$ with large vertical momentum flux $\langle v^{\prime }w^{\prime }\rangle$ . Filament arrest is induced by a lateral shear instability that generates horizontal momentum flux $\langle u^{\prime }v^{\prime }\rangle$ at low wavenumbers. The turbulent vertical velocity patterns, indicative of coherent Langmuir cells, change markedly across the horizontal domain with both across-filament and down-filament winds under the action of submesoscale currents.

Mechanics↗

The effect of filament peaking on electron beam weld quality

Electron beam welding uses high intensity beams that are generated by an electron emitting cathode under high vacuum conditions. The cathode, typically a ribbon or wire filament of refractory metal, must be heated to sufficiently high temperatures for electron emission to occur and must be carefully controlled to produce the desired beam current while at the same time optimizing filament lifetime. Traditional methods for doing this in commonly used triode gun assemblies consist of first finding the filament knee point in the filament current versus beam current relationship that produces the desired beam current for a given beam voltage. The filament current is then increased by some amount, typically 5–10%, per conventional wisdom to produce a peaked beam. This investigation studies and quantifies the beam and weld quality produced by underkneed and overpeaked filaments using electron beam diagnostics to measure the power density distribution of these beams. Underkneed filaments are shown to have power densities that drop off very quickly below the traditional knee, resulting in poor weld penetration. Kneed filaments do not reach the full beam intensity and also produce shallow welds. Increasing the filament current 10% above the traditional knee was shown to produce a circular Gaussian-like beam with peaked intensity optimized for welding and for filament lifetime. Further increases in the filament current that produce overpeaked beams do not significantly change the beam or weld quality and would only contribute to a reduced filament lifetime.

Electron Beam Welding↗

High-resolution Observations of Sympathetic Filament Eruptions by NVST

We investigate two sympathetic filament eruptions observed by the New Vacuum Solar Telescope on 2015 October 15. The full picture of the eruptions is obtained from the corresponding Solar Dynamics Observatory ( SDO )/Atmospheric Imaging Assembly (AIA) observations. The two filaments start from active region NOAA 12434 in the north and end in one large quiescent filament channel in the south. The left filament erupts first, followed by the right filament eruption about 10 minutes later. Clear twist structure and rotating motion are observed in both filaments during the eruption. Both eruptions failed, since the filaments first rise up, then flow toward the south and merge into the southern large quiescent filament. We also observe repeated activations of mini filaments below the right filament after its eruption. Using magnetic field models constructed based on SDO /HMI magnetograms via the flux rope insertion method, we find that the left filament eruption is likely to be triggered by kink instability, while the weakening of overlying magnetic fields due to magnetic reconnection at an X-point between the two filament systems might play an important role in the onset of the right filament eruption.

79 ASTRONOMY AND ASTROPHYSICS↗