Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein

The serine/threonine kinases ROCK1 and ROCK2 are central mediators of actomyosin contractile force generation that act downstream of the RhoA small GTP-binding protein. As a result, they have key roles in regulating cell morphology and proliferation, and have been implicated in numerous pathological conditions and diseases including hypertension and cancer. Here we describe the generation of a gene-targeted mouse line that enables CRE-inducible expression of a conditionally-active fusion between the ROCK2 kinase domain and the hormone-binding domain of a mutated estrogen receptor (ROCK2:ER). This two-stage system of regulation allows for tissue-selective expression of the ROCK2:ER fusion protein, which then requires administration of estrogen analogues such as tamoxifen or 4-hydroxytamoxifen to elicit kinase activity. This conditional gain-of-function system was validated in multiple tissues by crossing with mice expressing CRE recombinase under the transcriptional control of cytokeratin14 (K14), murine mammary tumor virus (MMTV) or cytochrome P450 Cyp1A1 (Ah) promoters, driving appropriate expression in the epidermis, mammary or intestinal epithelia respectively. Given the interest in ROCK signaling in normal physiology and disease, this mouse line will facilitate research into the consequences of ROCK activation that could be used to complement conditional knockout models

Precision control of thermal transport in cryogenic single-crystal silicon devices

We report on the diffusive-ballistic thermal conductance of multi-moded single-crystal silicon beams measured below 1 K. It is shown that the phonon mean-free-path $\ell$ is a strong function of the surface roughness characteristics of the beams. This effect is enhanced in diffuse beams with lengths much larger than $\ell$, even when the surface is fairly smooth, 5-10 nm rms, and the peak thermal wavelength is 0.6 $\mu$m. Resonant phonon scattering has been observed in beams with a pitted surface morphology and characteristic pit depth of 30 nm. Hence, if the surface roughness is not adequately controlled, the thermal conductance can vary significantly for diffuse beams fabricated across a wafer. In contrast, when the beam length is of order $\ell$, the conductance is dominated by ballistic transport and is effectively set by the beam area. We have demonstrated a uniformity of $\pm$8% in fractional deviation for ballistic beams, and this deviation is largely set by the thermal conductance of diffuse beams that support the micro-electro-mechanical device and electrical leads. In addition, we have found no evidence for excess specific heat in single-crystal silicon membranes. This allows for the precise control of the device heat capacity with normal metal films. We discuss the results in the context of the design and fabrication of large-format arrays of far-infrared and millimeter wavelength cryogenic detectors

Context and Political Knowledge: Explaining Cross-National Variation in Partisan Left-Right Knowledge

We present a theory that links variation in aggregate levels of political knowledge across countries and over time to corresponding differences in the political context in which voters become (or do not become) informed. Specifically, we argue that the level of partisan left-right knowledge in a given context ultimately depends on how useful the left-right metaphor is for organizing, simplifying, or otherwise facilitating voters’ understanding of political processes. Using survey data on the distribution of left-right knowledge in 59 different contexts (in 18 countries), our analysis reveals that voters understand the relative left-right positioning of parties to a much greater degree when these positions are important predictors of the composition of policy-making coalitions, but that variation in this knowledge does not correspond to the accuracy with which the relative left-right positions of parties predicts more narrow policy positions

Nonperturbative Effects from the Resummation of Perturbation Theory

Using the general argument in Borel resummation of perturbation theory that links the divergent perturbation theory to the nonperturbative effect we argue that the nonperturbative effect associated with the perturbation theory should have a branch cut only along the positive real axis in the complex coupling plane. The component in the weak coupling expansion of the nonperturbative amplitude, which usually includes the leading term in the weak coupling expansion, that gives rise to the branch cut can be calculated in principle from the perturbation theory combined with some exactly calculable properties of the nonperturbative effect. The realization of this mechanism is demonstrated in the double well potential and the two-dimensional O(N) nonlinear sigma model. In these models the leading term in weak coupling of the nonperturbative effect can be obtained with good accuracy from the first terms of the perturbation theory. Applying this mechanism to the infrared renormalon induced nonperturbative effect in QCD, we suggest some of the QCD condensate effects can be calculated in principle from the perturbation theory.Comment: 21 Pages, 1 Figure; To appear in Phys Rev

Design, Fabrication, and Testing of Lumped Element Kinetic inductance Detectors for 3 mm CMB Observations

Kinetic inductance detectors (KIDs) are a promising technology for low-noise, highly-multiplexible mm- and submm-wave detection. KIDs have a number of advantages over other detector technologies, which make them an appealing option in the cosmic microwave background B-mode anisotropy search, including passive frequency domain multiplexing and relatively simple fabrication, but have suffered from challenges associated with noise control. Here we describe design and fabrication of a 20-pixel prototype array of lumped element molybdenum KIDs. We show Q, frequency and temperature measurements from the array under dark conditions. We also present evidence for a double superconducting gap in molybdenum

Assessment of wind speeds along the damage path of the Alonsa, Manitoba EF4 tornado on 3 August 2018

Given the impracticality of attempting to directly measure wind speeds in tornadoes, wind speed estimation typically relies on the assessment of damage to structures and vegetation using classifications described in the Enhanced Fujita (EF) Scale. The advent of technology enabling the collection of large amounts of data, including detailed ground, drone, and aerial imagery, has led to a growth in research on non-conventional approaches for estimating tornado wind speeds. Research methods focused on defining the tornadic wind field based on forensic analysis of damage observations have shown promise for improving tornado assessments in a quantitative manner. In this study, novel methods for collecting forensic data following tornadoes are presented. Data from the Alonsa, MB tornado are applied to estimating the wind field along the damage path using treefall pattern analysis and threshold debris flight speed calculations. Comparison of the resulting wind speed estimates show reasonable agreement, with maximum speeds from both methods in the EF5 range. These research methods yield higher wind speeds than the maximum value obtained from the conventional EF-Scale assessment, which is in the low-end of the EF4 range based on a wood-frame house with sub-standard construction that was swept entirely from its foundation. Further work is still needed to make these methods operational for routine tornado intensity estimates

Scale Setting in QCD and the Momentum Flow in Feynman Diagrams

We present a formalism to evaluate QCD diagrams with a single virtual gluon using a running coupling constant at the vertices. This method, which corresponds to an all-order resummation of certain terms in a perturbative series, provides a description of the momentum flow through the gluon propagator. It can be viewed as a generalization of the scale-setting prescription of Brodsky, Lepage and Mackenzie to all orders in perturbation theory. In particular, the approach can be used to investigate why in some cases the ``typical'' momenta in a loop diagram are different from the ``natural'' scale of the process. It offers an intuitive understanding of the appearance of infrared renormalons in perturbation theory and their connection to the rate of convergence of a perturbative series. Moreover, it allows one to separate short- and long-distance contributions by introducing a hard factorization scale. Several applications to one- and two-scale problems are discussed in detail.Comment: eqs.(51) and (83) corrected, minor typographic changes mad

Explorations, Vol. 2, No. 2

Cover: The painting reproduced on the cover is an oil on canvas entitled “Konrad Oberhuber’s Visit to Compass Harbor, Maine, ” by Michael Lewis, 1985. Lewis is Professor of Art at the University of Maine at Orono. ©Michael Lewis, 1985 Articles include: The Gulf of Maine: A Sea Beside a Sea, by Kathleen Lignell The Gulf of Maine Littoral world of promise, by Carole J. Bombard for David Sanger Marine Worms Worth Fifteen Dollars Each? by David Dean Free Trade, Not Trade War, by James A. Wilson Inner Space—The Gulf of Maine: its history and future for research, by Robert S. Steneck The Shape of Mud and Its Importance to Marine Animals, by Les Watling The Humble Herring gold-plated sardine, by David K. Stevenson Seismic Profiling: The Search for the Submerged Geological Record in the Gulf of Maine, by Daniel F. Belknap and Joseph T. Kelley Our Cover Artist: Michael Lewis In the Classroom, by Michael Brody Nobody Told the Bumblebee He Couldn\u27t Fly, by Herbert Hidu The Tie That Binds: technology and research, by Robert S. Steneck Diet Developments for the Maine Lobster, by Robert Bayer A Wet Desert: secrets of the salt marsh, by Gary M. King Maine Marshes, by George L. Jacobson Mya Arenaria: return of the clam, by Devon Phillips CES: commitment and action, by Devon Phillips EPSCoR benthic research, by Lawrence M. Mayer In the Field of Researc

Pseudomonas aeruginosa arylsulfatase: a purified enzyme for the mild hydrolysis of steroid sulfates

The hydrolysis of sulfate ester conjugates is frequently required prior to analysis for a range of analytical techniques including gas chromatography-mass spectrometry (GC-MS). Sulfate hydrolysis may be achieved with commercial crude arylsulfatase enzyme preparations such as that derived from Helix pomatia but these contain additional enzyme activities such as glucuronidase, oxidase, and reductase that make them unsuitable for many analytical applications. Strong acid can also be used to hydrolyze sulfate esters but this can lead to analyte degradation or increased matrix interference. In this work, the heterologously expressed and purified arylsulfatase from Pseudomonas aeruginosa is shown to promote the mild enzyme-catalyzed hydrolysis of a range of steroid sulfates. The substrate scope of this P. aeruginosa arylsulfatase hydrolysis is compared with commercial crude enzyme preparations such as that derived from H. pomatia. A detailed kinetic comparison is reported for selected examples. Hydrolysis in a urine matrix is demonstrated for dehydroepiandrosterone 3-sulfate and epiandrosterone 3-sulfate. The purified P. aeruginosa arylsulfatase contains only sulfatase activity allowing for the selective hydrolysis of sulfate esters in the presence of glucuronide conjugates as demonstrated in the short three-step chemoenzymatic synthesis of 5α-androstane-3β,17β-diol 17-glucuronide (ADG, 1) from epiandrosterone 3-sulfate. The P. aeruginosa arylsulfatase is readily expressed and purified (0.9 g per L of culture) and thus provides a new and selective method for the hydrolysis of steroid sulfate esters in analytical sample preparation.We thank the Australian Government Anti-Doping Research Pro-gram for financial support