Native Mass Spectrometry for the Study of PROTAC GNE-987-Containing Ternary Complexes

PRO teolysis TA rgeting C himeras (PROTACs) promote the degradation, rather than inhibition, of a drug target as a mechanism for therapeutic treatment. Bifunctional PROTAC molecules allow simultaneous binding of both the target protein and an E3-Ubiquitin ligase, bringing the two proteins into close spatial proximity to allow ubiquitinylation and degradation of the target protein via the cell's endogenous protein degradation pathway. We utilized native mass spectrometry (MS) to study the ternary complexes promoted by the previously reported PROTAC GNE-987 between Brd4 bromodomains 1 and 2, and Von Hippel Lindeau E3-Ubiquitin Ligase. Native MS at high resolution allowed us to measure ternary complex formation as a function of PROTAC concentration to provide a measure of complex affinity and stability, whilst simultaneously measuring other intermediate protein species. Native MS provides a high-throughput, low sample consumption, direct screening method to measure ternary complexes for PROTAC development.     

Development and evaluation of an ultrasonic personal aerosol sampler

Assessing personal exposure to air pollution has long proven challenging due to technological limitations posed by the samplers themselves. Historically, wearable aerosol monitors have proven to be expensive, noisy, and burdensome. The objective of this work was to develop a new type of wearable monitor, an ultrasonic personal aerosol sampler (UPAS), to overcome many of the technological limitations in personal exposure assessment. The UPAS is a time‐integrated monitor that features a novel micropump that is virtually silent during operation. A suite of onboard environmental sensors integrated with this pump measure and record mass airflow (0.5–3.0 L/min, accurate within 5%), temperature, pressure, relative humidity, light intensity, and acceleration. Rapid development of the UPAS was made possible through recent advances in low‐cost electronics, open‐source programming platforms, and additive manufacturing for rapid prototyping. Interchangeable cyclone inlets provided a close match to the EPA PM_2.5 mass criterion (within 5%) for device flows at either 1.0 or 2.0 L/min. Battery life varied from 23 to 45 hours depending on sample flow rate and selected filter media. Laboratory tests of the UPAS prototype demonstrate excellent agreement with equivalent federal reference method samplers for gravimetric analysis of PM_2.5 across a broad range of concentrations.     

Predictors of summertime heat index levels in New York City apartments

During heat waves, fatal overexposure to heat most often occurs at home. It is not known how factors such as building size, floor level, and different types of air conditioning (AC) contribute to excess indoor heat. We monitored indoor temperature and humidity in 36 apartments in New York City during summers 2014 and 2015 and used these values to calculate the indoor heat index (HI). We investigated the role of AC type and building‐level factors on indoor HI using multilevel regression models. Thirty‐four of 36 homes had AC. Central and ductless AC types were associated with the coolest indoor conditions; homes with window and portable AC were significantly warmer. Apartments on the top floor of a building were significantly hotter during heat advisory periods than other apartments regardless of the presence of AC. High indoor HI levels persisted in some homes for approximately 1 day following the end of the two heat advisory periods. We provide concrete evidence of higher heat levels in top floor apartments and in homes with certain types of AC. High heat levels that persist indoors after outdoor heat has subsided may present an underappreciated public health risk.     

The coalescence of H2S and CO2 bubbles in water

Coalescence times are reported for bubbles of H₂S and CO₂ formed on adjacent nozzles in water. With CO₂, times were small (1–3 ms) at pressures below 2 MPa, and rose to 20 ms at 3.4 MPa. With H₂S, the logarithm of the coalescence time was proportional to pressure, and coalescence times reached 100 ms at 1.7 MPa. Film stretching rates were calculated assuming that the film surfaces were slightly immobile. By using a form of Vrij's theory for the inherent breaking time of films of constant thickness, the effective number of immobile layers could be calculated. Since these were in the range 10^?2 to 10^?3, only a small amount of “stiffness” in the film surface is enough to cause large changes in coalescence time.     

The value of a desk study for building a national river obstacle inventory

This study evaluates two desk‐based approaches for building an inventory of man‐made river obstacles. The creation of a river obstacle inventory is a vital first step in developing a prioritization process for obstacle removal and/or modification. In this study, a desktop geographical information system analysis of two rivers and their tributary network was undertaken, using two different approaches. The first involved analysing historical maps, satellite imagery, and Ordnance Survey Ireland Discovery Series maps and producing a geo‐referenced layer of all the potential river obstacles. The second involved developing a geo‐referenced layer of potential river obstacles based on the intersections between elements of the transport network (roads and railways) and river systems. To determine the effectiveness of the desk studies, the located obstacles were cross‐referenced with actual obstacles verified through a field survey. The desk studies identified several thousand potential obstacles. The study utilizing a range of maps consistently located a greater number of actual obstacles than the desk study based on intersections between the transport and river networks. The results indicate that desk‐based research offers an efficient and effective method for locating river obstacles and can guide subsequent field surveys aimed at confirming the presence of obstacles. This is particularly useful for eliminating from study large stretches of rivers that would otherwise need to be walked to confirm the presence, or otherwise, of potential river obstacles. In this regard, desk‐based exercises can offer opportunities to save on both time and cost in larger river assessments.     

The impacts of cattle access points on deposited sediment levels in headwater streams in Ireland

Cattle access to streams has been linked globally with degradation of stream water quality, driven largely by bank erosion and resultant instream, fine sediment deposition. The majority of evidence on such effects is however based in arid and semiarid regions of the United States and Australia, with few studies relating to cool temperate climates such as Northwest Europe. In this study, “Quorer” resuspendable sediment samples were taken from riffle geomorphic units upstream (control) and at two points downstream (pressure and recovery) of cattle access points in headwater streams in agricultural catchments in Ireland to assess levels of deposited stream sediment. Samples were taken in April/May (2016) prior to the grazing season and in October (2016) at the end of the grazing season. Sites in good‐high ecological status catchments and less than good ecological status catchments were included in the study. Higher levels of sediment were found downstream of cattle access points in both good‐high status and less than good status catchments; however, the impacts of access points were spatially confined to, in most cases, the area immediately downstream of the point of access. There was a strong correlation between deposited sediment mass and organic matter (OM) mass, with levels of OM increasing linearly with deposited sediment mass. Levels of measured sediment were negatively correlated with riparian habitat health (measured using a qualitative habitat assessment). The results of this study highlight the need for measures to prevent cattle access to headwater streams where access points can be many in order to manage local habitat quality and downstream water quality issues.