Engineering topics

Broughton, Howard

Publications and source records attributed to Broughton, Howard.

7 records

DOE OSTI2022

Crystal structure of active CDK4-cyclin D and mechanistic basis for abemaciclib efficacy

Despite the biological and therapeutic relevance of CDK4/6 for the treatment of HR+, HER2- advanced breast cancer, the detailed mode of action of CDK4/6 inhibitors is not completely understood. Of particular interest, phosphorylation of CDK4 at T172 (pT172) is critical for generating the active conformation, yet no such crystal structure has been reported to date. We describe here the x-ray structure of active CDK4-cyclin D3 bound to the CDK4/6 inhibitor abemaciclib and discuss the key aspects of the catalytically-competent complex. Furthermore, the effect of CDK4/6 inhibitors on CDK4 T172 phosphorylation has not been explored, despite its role as a potential biomarker of CDK4/6 inhibitor response. We show mechanistically that CDK4/6i stabilize primed (pT172) CDK4-cyclin D complex and selectively displace p21 in responsive tumor cells. Stabilization of active CDK4-cyclin D1 complex can lead to pathway reactivation following alternate dosing regimen. Consequently, sustained binding of abemaciclib to CDK4 leads to potent cell cycle inhibition in breast cancer cell lines and prevents rebound activation of downstream signaling. Overall, our study provides key insights demonstrating that prolonged treatment with CDK4/6 inhibitors and composition of the CDK4/6-cyclin D complex are both critical determinants of abemaciclib efficacy, with implications for this class of anticancer therapy.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2020

Integration of phage and yeast display platforms: A reliable and cost effective approach for binning of peptides as displayed on-phage

Hundreds of target specific peptides are routinely discovered by peptide display platforms. However, due to the high cost of peptide synthesis only a limited number of peptides are chemically made for further analysis. Here we describe an accurate and cost effective method to bin peptides on-phage based on binding region(s), without any requirement for peptide or protein synthesis. This approach, which integrates phage and yeast display platforms, requires display of target and its alanine variants on yeast. Flow cytometry was used to detect binding of peptides on-phage to the target on yeast. Once hits were identified, they were synthesized to confirm their binding region(s) by HDX (Hydrogen deuterium exchange) and crystallography. Moreover, we have successfully shown that this approach can be implemented as part of a panning process to deplete non-functional peptides. This technique can be applied to any target that can be successfully displayed on yeast; it narrows down the number of peptides requiring synthesis; and its utilization during selection results in enrichment of peptide population against defined binding regions on the target.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2005

Local and Total Density Measurements in Ice Shapes

Preliminary measurements of local and total densities inside ice shapes were obtained from ice shapes grown in the NASA Glenn Research Tunnel for a range of glaze ice, rime ice, and mixed phase ice conditions on a NACA 0012 airfoil at 0 angle of attack. The ice shapes were removed from the airfoil and a slice of ice 3 mm thick was obtained using a microtome. The resulting samples were then x-rayed to obtain a micro-radiography, the film was digitized, and image processing techniques were used to extract the local and total density values.

Vargas, Mario↗

NASA NTRS1999

Video Imaging System Particularly Suited for Dynamic Gear Inspection

A digital video imaging system that captures the image of a single tooth of interest of a rotating gear is disclosed. The video imaging system detects the complete rotation of the gear and divide that rotation into discrete time intervals so that each tooth of interest of the gear is precisely determined when it is at a desired location that is illuminated in unison with a digital video camera so as to record a single digital image for each tooth. The digital images are available to provide instantaneous analysis of the tooth of interest, or to be stored and later provide images that yield a history that may be used to predict gear failure, such as gear fatigue. The imaging system is completely automated by a controlling program so that it may run for several days acquiring images without supervision from the user.

Broughton, Howard↗

NASA NTRS1996

Validation of NASA Thermal Ice Protection Computer Codes: Program Overview - Part 1

The Icing Technology Branch at NASA Lewis has been involved in an effort to validate two thermal ice protection codes developed at the NASA Lewis Research Center. LEWICE/Thermal (electrothermal deicing & anti-icing), and ANTICE (hot-gas & electrothermal anti-icing). The Thermal Code Validation effort was designated as a priority during a 1994 'peer review' of the NASA Lewis Icing program, and was implemented as a cooperative effort with industry. During April 1996, the first of a series of experimental validation tests was conducted in the NASA Lewis Icing Research Tunnel(IRT). The purpose of the April 96 test was to validate the electrothermal predictive capabilities of both LEWICE/Thermal, and ANTICE. A heavily instrumented test article was designed and fabricated for this test, with the capability of simulating electrothermal de-icing and anti-icing modes of operation. Thermal measurements were then obtained over a range of test conditions, for comparison with analytical predictions. This paper will present an overview of the test, including a detailed description of: (1) the validation process; (2) test article design; (3) test matrix development; and (4) test procedures. Selected experimental results will be presented for de-icing and anti-icing modes of operation. Finally, the status of the validation effort at this point will be summarized. Detailed comparisons between analytical predictions and experimental results are contained in the following two papers: 'Validation of NASA Thermal Ice Protection Computer Codes: Part 2- The Validation of LEWICE/Thermal' and 'Validation of NASA Thermal Ice Protection Computer Codes: Part 3-The Validation of ANTICE'

Miller, Dean↗

NASA NTRS1996

Determination of Shed Ice Particle Size Using High Speed Digital Imaging

A full scale model of an aircraft engine inlet was tested at NASA Lewis Research Center's Icing Research Tunnel. Simulated natural ice sheds from the engine inlet lip were studied using high speed digital image acquisition and image analysis. Strategic camera placement integrated at the model design phase allowed the study of ice accretion on the inlet lip and the resulting shed ice particles at the aerodynamic interface plane at the rear of the inlet prior to engine ingestion. The resulting digital images were analyzed using commercial and proprietary software to determine the size of the ice particles that could potentially be ingested by the engine during a natural shedding event. A methodology was developed to calibrate the imaging system and insure consistent and accurate measurements of the ice particles for a wide range of icing conditions.

Broughton, Howard↗

NASA NTRS1996

Determination of Local Densities in Accreted Ice Samples Using X-Rays and Digital Imaging

At the NASA Lewis Research Center's Icing Research Tunnel ice shapes, similar to those which develop in-flight icing conditions, were formed on an airfoil. Under cold room conditions these experimental samples were carefully removed from the airfoil, sliced into thin sections, and x-rayed. The resulting microradiographs were developed and the film digitized using a high resolution scanner to extract fine detail in the radiographs. A procedure was devised to calibrate the scanner and to maintain repeatability during the experiment. The techniques of image acquisition and analysis provide accurate local density measurements and reveal the internal characteristics of the accreted ice with greater detail. This paper will discuss the methodology by which these samples were prepared with emphasis on the digital imaging techniques.

Broughton, Howard↗