Abbottgodwin7479

All rights reserved. For permissions, please e-mail journals.permissions@oup.com.Minority Gene Expression Profiling (MGEP) refers to a scenario where the expression profiles of specific genes of interest are concentrated in a small cellular pool that is embedded within a larger, non-expressive pool. An example of this is the analysis of disease-related genes within sub-populations of blood or biopsied tissues. These systems are characterized by low signal-to-noise ratios that make it difficult, if not impossible, to uncover the desired signatures of pathogenesis in the absence of lengthy, and often problematic, technical manipulations. We have adapted ribosome profiling (RP) workflows from the Illumina to the Ion Proton platform and used them to analyze signatures of pathogenesis in an MGEP model system consisting of human cells eliciting less then 3% productive dengue infection. We find that RP is powerful enough to identify relevant responses of differentially expressed genes, even in the presence of significant noise. We discuss how to deal with sources of unwanted variation, and propose ways to further improve this powerful approach to the study of pathogenic signatures within MGEP systems. © The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.Preventing the transfer of allergens from one food to another via food-contact surfaces in retail food environments is an important aspect of retail food safety. Existing recommendations for wiping and cleaning food-contact surfaces is mainly focused on preventing microorganisms such as bacteria and viruses from contaminating foods. The effectiveness of these wiping and cleaning recommendations for preventing the transfer of food allergens in retail and food service establishments remains unclear. This project investigated 1) allergen removal from surfaces by wiping with paper wipes, terry cloths and alcohol/quaternary ammonium chloride (quat) sanitizing wipes; 2) cleaning of allergen-contaminated surfaces using a wash-rinse-sanitize-air dry procedure; and 3) allergen transfer from contaminated wipes to multiple surfaces. Food-contact surfaces (stainless steel, textured plastic and maple wood) were contaminated with peanut-, milk- and egg-containing foods, and subjected to various wiping and cleaning procedures. For transfer experiments, dry paper wipes or wet cloths contaminated with allergenic foods were wiped on four surfaces of the same composition. Allergen-specific lateral flow devices were used to detect the presence of allergen residues on wiped or cleaned surfaces. While dry wipes and cloths were not effective for removing allergenic foods, terry cloths pre-soaked in water or sanitizer solution, use of multiple quat wipes, and the wash-rinse-sanitize-air dry procedure were effective in allergen removal from surfaces. Allergens present on dry wipes were transferred to wiped surfaces. In contrast, minimal or no allergen transfer to surfaces was found when allergen-contaminated terry cloths were submerged in sanitizer solution prior to wiping surfaces. The full cleaning method (wash-rinse-sanitize-air dry) and soaking the terry cloth in sanitizer solution prior to wiping were effective at allergen removal and minimizing allergen transfer.Large-conductance Ca2+-activated K+ channels (BK channels) are activated by cytosolic calcium and depolarized membrane potential under physiological conditions. Thus, these channels control electrical excitability in neurons and smooth muscle by gating K+ efflux and hyperpolarizing the membrane in response to Ca2+ signaling. Opaganib Altered BK channel function has been linked to epilepsy, dyskinesia, and other neurological deficits in humans, making these channels a key target for drug therapies. To gain insight into mechanisms underlying pharmacological modulation of BK channel gating, here we studied mechanisms underlying activation of BK channels by the biarylthiourea derivative, NS11021, which acts as a smooth muscle relaxant. We observe that increasing NS11021 shifts the half-maximal activation voltage for BK channels toward more hyperpolarized voltages, in both the presence and nominal absence of Ca2+, suggesting that NS11021 facilitates BK channel activation primarily by a mechanism that is distinct from Ca2+ activation. 30 µM NS11021 slows the time course of BK channel deactivation at -200 mV by ∼10-fold compared with 0 µM NS11021, while having little effect on the time course of activation. This action is most pronounced at negative voltages, at which the BK channel voltage sensors are at rest. Single-channel kinetic analysis further shows that 30 µM NS11021 increases open probability by 62-fold and increases mean open time from 0.15 to 0.52 ms in the nominal absence of Ca2+ at voltages less than -60 mV, conditions in which BK voltage sensors are largely in the resting state. We could therefore account for the major activating effects of NS11021 by a scheme in which the drug primarily shifts the pore-gate equilibrium toward the open state. © 2020 Rockman et al.Researchers identify key residue in GluN2A subunit that may regulate channel opening by organizing a network of aromatic amino acids. © 2020 Rockefeller University Press.The NMDA receptor (NMDAR) is an ionotropic glutamate receptor formed from the tetrameric assembly of GluN1 and GluN2 subunits. Within the flexible linker between the agonist binding domain (ABD) and the M1 helix of the pore-forming transmembrane helical bundle lies a two-turn, extracellular pre-M1 helix positioned parallel to the plasma membrane and in van der Waals contact with the M3 helix thought to constitute the channel gate. The pre-M1 helix is tethered to the bilobed ABD, where agonist-induced conformational changes initiate activation. Additionally, it is a locus for de novo mutations associated with neurological disorders, is near other disease-associated de novo sites within the transmembrane domain, and is a structural determinant of subunit-selective modulators. To investigate the role of the pre-M1 helix in channel gating, we performed scanning mutagenesis across the GluN2A pre-M1 helix and recorded whole-cell macroscopic and single channel currents from HEK293 cell-attached patches. We identified two residues at which mutations perturb channel open probability, the mean open time, and the glutamate deactivation time course.