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Analysis of strategies to meet ASHRAE S241 infectious aerosol control targets by space type and region using EnergyPlus™ simulations

Professional organizations, such as ASHRAE, have recently proposed new voluntary standards for controlling infectious aerosols. Specifically, ASHRAE Standard 241 (S241) defines equivalent clean air targets for a range of space types that can be met through combinations of mitigation measures. This paper seeks to inform the selection of measures by space type and climate zone through building simulations that quantify the impacts of increased outdoor air ventilation; increasing filtration and/or adding germicidal ultraviolet (GUV) in the central heating, ventilation, and air-conditioning (HVAC) systems; or using portable air cleaners (PACs), upper room GUV, or whole room GUV approaches to increase equivalent clean air delivery. The measures are assessed individually and in practical combinations for their ability to meet S241 in four space types (offices, classrooms, dining areas, and healthcare waiting rooms) using prototype buildings modeled using EnergyPlus. The mitigation measures are compared holistically against baseline building operations using metrics for equivalent clean air, energy, and comfort. The results in this study show that single measures can meet S241 for offices with minimal impacts on energy and comfort, while either upper room GUV systems or combinations of measures such as MERV 13 HVAC filtration with PACs are needed to meet S241 for classrooms. The dining area and healthcare waiting room targets cannot be met in this study when assuming design occupancy and MS2 as the challenge agent, even when combining multiple measures together. This paper provides valuable considerations when designing measures to meet S241 for a range of spaces and scenarios.

GUV

Protocol for Engineered Compositional Asymmetry Within Nanodiscs

Membrane proteins remain the most challenging targets for structural characterization, yet their elucidation provides valuable insights into protein function, disease mechanisms, and drug specificity. Structural biology platforms have advanced rapidly in recent years, notably through the development and implementation of nanodiscs—discoidal lipid–protein complexes that encapsulate and solubilize membrane proteins within a controlled, native-like environment. While nanodiscs have become powerful tools for studying membrane proteins, faithfully reconstituting the compositional asymmetry intrinsic to nearly all biological membranes has not yet been achieved. Proper membrane leaflet lipid distribution is critical for accurate protein folding, stability, and insertion. Here, we share a protocol for reconstituting tailored compositional asymmetry within nanodiscs through membrane extraction from giant unilamellar vesicles (GUVs) treated with a leaflet-specific methyl-β-cyclodextrin (mβCD) lipid exchange. Nanodisc asymmetry is verified through a geometric approach: biotin-DPPE-preloaded mβCD engages in lipid exchange with the outer leaflet of POPC GUVs solubilized by the lipid-free membrane scaffold protein (MSP) Δ49ApoA-I to form nanodisc structures. Once isolated, nanodiscs are introduced to the biotin-binding bacterial protein streptavidin. High-speed atomic force microscopy imaging depicts nanodisc–dimer complexes, indicating that biotin-DPPE was successfully reconstituted into a single leaflet of the nanodiscs. This finding outlines the first step toward engineering tailored nanodisc asymmetry and mimicking the native environment of integral proteins—a potentially powerful tool for accurately reconstituting and structurally analyzing integral membrane proteins whose functions are modulated by lipid asymmetry.

Biological and medical sciences