Clinically Applicable Assessment of Tisagenlecleucel CAR T Cell Treatment by Digital Droplet PCR for Copy Number Variant Assessment

Chimeric antigen receptor (CAR) T cell therapy is an innovative immunotherapy for treating cancers in both children and adults with proven utility in numerous clinical trials. Significantly, some CAR T cell therapies have now been approved by relevant national regulatory bodies across numerous countries for clinical therapeutic use outside of clinical trials. One such recently licensed product is tisagenlecleucel, a CAR T therapy approved for the treatment of B-cell acute lymphoblastic leukemia (B-ALL) using autologous T cells from the patient. The genetically engineered T cells target a protein called CD19, common to B cells, through a CAR incorporating a 4-1BB costimulatory domain to improve response. Since tisagenlecleucel is now a standard of care treatment for B-ALL, it is clinically essential to be able to accurately monitor these CAR T cells in patients. Assessment of the copy number variant (CNV) of the CAR T cell products allows this within a clinically acceptable timeframe for optimal patient benefit. However, no standardized method with high reproducibility and efficiency has been described within a routine clinical laboratory setting. Here, we demonstrated a novel digital droplet PCR (ddPCR)-based methodology for the study of CNV (ddPCR-CNV) in 4-1BB CD19-specific CAR T cells with universal applicability across clinical diagnostic laboratories.     

Electromagnetic Metrology on Concrete and Corrosion

To augment current methods for the evaluation of reinforcing bar (rebar) corrosion within concrete, we are exploring unique features in the dielectric and magnetic spectra of pure iron oxides and corrosion samples. Any signature needs to be both prominent and consistent in order to identify corrosion within concrete bridge deck or other structures. In order to measure the permittivity and propagation loss through concrete as a function of temperature and humidity, we cut and carefully fitted samples from residential concrete into three different waveguides. We also poured and cured a mortar sample within a waveguide that was later measured after curing 30 days. These measurements were performed from 45 MHz to 12 GHz. Our concrete measurements showed that the coarse granite aggregate that occupied about half the sample volume reduced the electromagnetic propagation loss in comparison to mortar. We also packed ground corrosion samples and commercially available iron-oxide powders into a transmission-line waveguide and found that magnetite and corrosion sample spectra are similar, with a feature between 0.5 GHz and 2 GHz that may prove useful for quantifying corrosion. We also performed reflection (S₁₁) measurements at various corrosion surfaces and in loose powders from 45 MHz to 50 GHz. These results are a first step towards quantifying rebar corrosion in concrete.     

Numerical experience with diagonalization algorithms for asymmetric demand traffic assignment

This paper reviews the concept of a diagonalization algorithm for use in solving traffic network equilibrium problems for which the arc cost and/or the origin-destination travel demand functions are asymmetric. Such functions are known to occur in realistic settings involving multiple modes or users. The computational performance of this algorithm for different degrees of travel demand asymmetry is then explored by a detailed numerical experiment since no previous results of this type have been reported. It is found that, through the use of progressive stopping tolerances, the impact of high degrees of travel demand function asymmetry on the computational burden associated with finding a traffic network equilibrium may be mitigated; in effect equilibrium problems with high degrees of demand asymmetry are little more difficult to solve than perfectly symmetric problems.     

Simulating sympathetic detonation using the hydrodynamic models and constitutive equations

A Sympathetic detonation (SD) is a detonation of an explosive charge by a nearby explosion. Most of times it is unintended while the impact of blast fragments or strong shock waves from the initiating donor explosive is the cause of SD. We investigate the SD of a cylindrical explosive charge (64 % RDX, 20 % Al, 16 % HTPB) contained in a steel casing. The constitutive relations for high explosive are obtained from a thermo-chemical code that provides the size effect data without the rate stick data typically used for building the rate law and equation of state. A full size SD test of eight pallet-packaged artillery shells is performed that provides the pressure data while the hydrodynamic model with proper constitutive relations for reactive materials and the fragmentation model for steel casing is conducted to replicate the experimental findings. The work presents a novel effort to accurately model and reproduce the sympathetic detonation event with a reduced experimental effort.     

Process development for the laser powder bed fusion of WC-Ni Cermets using sintered-agglomerated powder

Although ceramic particle-metal matrix materials (i.e., cermets) can offer superior performance, manufacturing these materials via conventional means is difficult compared to the manufacturing of metal alloys. This study leverages the laser powder bed fusion (LPBF) process to additively manufacture dense tungsten carbide (WC)-17 wt.% nickel (Ni) composite specimens using novel spherical, sintered-agglomerated composite powder. A range of processing parameters yielding high-density specimens was discovered using a sequential series of experiments comprised of single bead, multi-layer, and cylindrical builds. Cylinders with a relative density >99% were fabricated and characterized in terms of microstructure, chemical composition, and hardness. Scanning electron microscopy images show favorable wetting between the Ni binder and carbide particles without any phase segregation and laser processing increased the average carbide particle size. Energy dispersive X-ray and X-ray diffraction analyses detected traces of secondary products after laser processing. For samples processed at high energy densities, complex carbides and carbon agglomerate phases were detected. The maximum hardness of 60.38 Rockwell C is achieved in the printed samples. The successful builds in this study open the way for LPBF of dense WC-Ni parts with a large workable laser power-laser velocity processing window.     

Investigation of the Sharkskin melt instability using optical Fourier analysis

An optical method allowing the characterization of melt flow instabilities typically occurring during an extrusion process of polymers and polymer compounds is presented. It is based on a camera-acquired image of the extruded compound with a reference length scale. Application of image processing and transformation of the calibrated image to the frequency domain yields the magnitude spectrum of the instability. The effectiveness of the before mentioned approach is shown on Styrene-butadiene rubber (SBR) compounds, covering a wide range of silica filler content, extruded through a Göttfert capillary rheometer. The results of the image-based analysis are compared with the results from the sharkskin option, a series of highly sensitive pressure transducers installed inside the rheometer. A simplified version of the code used to produce the optical analysis results is included as supplementary material. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48806.     

Silicon-29 solid-state nuclear magnetic resonance spectroscopy of composite interfaces

The types of structures and bonds that are formed with silicons in the composite interface were studied using ²⁹Si cross-polarization/magic angle sample spinning (CP/MAS) nuclear magnetic resonance spectroscopy (NMR). The change in mobility of silane coupling agent bonded to silica, as compared with bulk hydrolyzed silane coupling agent, can be monitored by the change in line width and the shift of resonances to higher fields, as well as by the change in the silicon-proton cross-polarization time T_SiH. In the silane coupling agent-matrix resin interface, the T_SiH values reflect the change in mobility as a function of the concentration and degree of hydrolysis of the silane coupling agent. It has been demonstrated that quantitative measurements of T_SiH can be used to investigate relative mobilities.