High-content screening and profiling of drug activity in an automated centrosome-duplication assay

Maintenance of centrosome number is essential for cell-cycle progression and genomic stability, but investigation of this regulation has been limited by assay difficulty. We present a fully automated image-based centrosome-duplication assay that is accurate and robust enough for both careful cell-biology studies and high-throughput screening, and employ this assay in a series of chemical-genetic studies. We observe that a simple cytometric profiling strategy, which is based on organelle size, groups compounds with similar mechanisms of action; this suggests a simple strategy for excluding compounds that undesirably target such activities as protein synthesis and microtubule dynamics. Screening a library of compounds of known activity, we found unexpected effects on centrosome duplication by a number of drugs, most notably isoform-specific protein kinase C inhibitors and retinoic acid receptor agonists. From a 16 320-member library of uncharacterized small molecules, we identified five potent centrosome-duplication inhibitors that do not target microtubule dynamics or protein synthesis. The analysis methodology reported here is directly relevant to studies of centrosome regulation in a variety of systems and is adaptable to a wide range of other biological problems.     

Quantitative use of Rayleigh waves to locate and size subsurface holes

An ultrasonic inspection method is used to obtain the circumference of a subsurface hole and the depth of the hole below the surface. A pitch-catch Rayleigh wave transducer set-up was used to launch a Rayleigh surface wave at the flaw and to capture and record the scattered waves. The frequency spectrum of the scattered waves can be used to obtain the depth of the hole. The ligament of material between the hole and the surface is sent into resonance, and this feature can be extracted from the scattered waves' frequency spectrum. The frequency is a function of the ligament length; thus the hole depth can be obtained. The circumference of the hole is found from a time of flight measurement. A Rayleigh wave is formed that travels around the hole's surface. The length of time required for the wave to travel around the hole is a measure of the circumference.     

Estimating the quantity of egg white and whey protein concentrate in prepared crabstick using ELISA

Indirect enzyme-linked immunosorbent assays (ELISA) technique was used to identify and quantify the use of dried egg white (DEW) and whey protein concentrate (WPC) in crabsticks. The use of SDS–PAGE for the quantification of protein additives has had limited success due to the high shear and high temperature processes of surimi crabstick. Monoclonal (anti-heat-denatured ovalbumin) and polyclonal (anti-β-lactoglobulin) antibodies were used. Antibodies showed no significant cross-reactivity with non-target crabstick proteins. An optimised extraction solution of 10% SDS and 2.5% 2-ME yielded high extractability with improved consistency. Quantification of DEW and WPC was achieved using the optimised extraction solution and indirect ELISA. Estimated DEW values were within 7% of actual values, WPC samples were within 17%. Inter-assay coefficients of variance for DEW ranged from 0.9% to 3.1% and those of the WPC were 1.0–8.0%.     

Afterburn Ignition Delay and Shock Augmentation in Fuel Rich Solid Explosives

We present experimental and computational results that explain some aspects of measured energy release in explosions of unconfined trinitrotoluene [TNT, C₆H₂(NO₂)₃CH₃], and an aluminum‐containing explosive formulation, and show how this energy release can influence shock wave velocities in air. In our interpretation, energy release is divided into early, middle, and late time regimes. An explanation is provided for the interdependence of the time regimes and their influence on the rate at which energy (detonation/explosion and afterburn) is released. We use a merging of the thermodynamic and chemical kinetic processes that predicts how chemical kinetics may determine the time delay of the afterburn of combustible gases produced by the initial detonation/explosion/fast reaction. The thermodynamic computer code CHEETAH is used to predict gaseous and solid products of early time energy release, and a chemical kinetic reaction mechanism (CHEMKIN format) is used to describe the subsequent afterburn of the gas phase products in air. Results of these calculations are compared with field measurements of unconfined explosions of 2 kg charge weights of TNT and an aluminum‐containing explosive formulation.     

The perfluoropolymer upper bound

Perfluoropolymers have fundamentally distinct thermodynamic partitioning properties compared to those of their hydrocarbon counterparts. However, current upper bound theory assumes hydrocarbon solubility behavior for all polymers. Herein, the fundamental presupposition of invariance in solubility behavior to upper bound performance is critically assessed for perfluoropolymers and hydrocarbon polymers. By modifying solubility relationships, theoretical perfluoropolymer upper bounds are established, showing a positive shift of the upper bound front as a result of beneficial solubility selectivities for certain gas pairs, including N₂/CH₄, He/H₂, He/N₂, He/CH₄, and He/CO₂. Within the framework of the solution–diffusion model, an analysis is presented to compare two independent approaches often pursued in efforts to surpass the polymer upper bound: (a) achieving solubility selectivity via perfluoropolymers and (b) improving diffusion selectivity via rigid hydrocarbon polymers. This analysis demonstrates the significant benefit that can be achieved by considering both the chemical composition and morphology of solid-state macromolecules when designing membrane materials.     

Oxygen-Generating Cryogels Restore T Cell Mediated Cytotoxicity in Hypoxic Tumors

Solid tumors are protected from antitumor immune responses due to their hypoxic microenvironments. Weakening hypoxia-driven immunosuppression by hyperoxic breathing of 60% oxygen has shown to be effective in unleashing antitumor immune cells against solid tumors. However, efficacy of systemic oxygenation is limited against solid tumors outside of lungs and has been associated with unwanted side effects. As a result, it is essential to develop targeted oxygenation alternatives to weaken tumor hypoxia as novel approaches to restore immune responses against cancer. Herein, injectable oxygen-generating cryogels (O₂-cryogels) to reverse tumor-induced hypoxia are reported. These macroporous biomaterials are designed to locally deliver oxygen, inhibit the expression of hypoxia-inducible genes in hypoxic melanoma cells, and reduce the accumulation of immunosuppressive extracellular adenosine. The data show that O₂-cryogels enhance T cell-mediated secretion of cytotoxic proteins, restoring the killing ability of tumor-specific cytotoxic T lymphocytes, both in vitro and in vivo. In summary, O₂-cryogels provide a unique and safe platform to supply oxygen as a coadjuvant in hypoxic tumors and have the potential to improve cancer immunotherapies.     

Magnetic measurements at pressures above 10 GPa in a miniature ceramic anvil cell for a superconducting quantum interference device magnetometer

A miniature ceramic anvil high pressure cell (mCAC) was earlier designed by us for magnetic measurements at pressures up to 7.6 GPa in a commercial superconducting quantum interference magnetometer [N. Tateiwa et al., Rev. Sci. Instrum. 82, 053906 (2011)]. Here, we describe methods to generate pressures above 10 GPa in the mCAC. The efficiency of the pressure generation is sharply improved when the Cu-Be gasket is sufficiently preindented. The maximum pressure for the 0.6 mm culet anvils is 12.6 GPa when the Cu-Be gasket is preindented from the initial thickness of 300-60 μm. The 0.5 mm culet anvils were also tested with a rhenium gasket. The maximum pressure attainable in the mCAC is about 13 GPa. The present cell was used to study YbCu(2)Si(2) which shows a pressure induced transition from the non-magnetic to magnetic phases at 8 GPa. We confirm a ferromagnetic transition from the dc magnetization measurement at high pressure. The mCAC can detect the ferromagnetic ordered state whose spontaneous magnetic moment is smaller than 1 μ(B) per unit cell. The high sensitivity for magnetic measurements in the mCAC may result from the simplicity of cell structure. The present study shows the availability of the mCAC for precise magnetic measurements at pressures above 10 GPa.     

The relationship between Algebra Nation usage and high‐stakes test performance for struggling students

Although the use of technology in the K12 classroom has been shown to have a positive impact, research on the use of open education resources (OER) is relatively limited, especially research focusing on low‐achieving students. The present study examines the relationship between usage of Algebra Nation, a self‐guided system that provided instructional videos and practice problems, and the performance of students who had failed the state‐administered Algebra I end‐of‐course (EOC) assessment the previous year. Indicators of usage of Algebra Nation consisted of logins, video views, and practice questions answered. Path analyses and logistic regressions were used to evaluate relationships between usage indicators and algebra scores, controlling for number of absences, free/reduced lunch eligibility, Hispanic/Latino origin, race, and gender. The results indicate that higher levels of logins, video views, and practice questions answered were related to higher scores when the students re‐took the assessment. Logins and practice questions were also related to increases in odds of passing the Algebra I EOC assessment, but not video views. The results suggest that there may be benefits to technology use in the form of an OER adopted by students and teachers on an informal basis and link self‐regulated learning strategies to student achievement.     

Lithium-Battery Anode Gains Additional Functionality for Neuromorphic Computing through Metal–Insulator Phase Separation

Specialized hardware for neural networks requires materials with tunable symmetry, retention, and speed at low power consumption. The study proposes lithium titanates, originally developed as Li-ion battery anode materials, as promising candidates for memristive-based neuromorphic computing hardware. By using ex- and in operando spectroscopy to monitor the lithium filling and emptying of structural positions during electrochemical measurements, the study also investigates the controlled formation of a metallic phase (Li₇Ti₅O₁₂) percolating through an insulating medium (Li₄Ti₅O₁₂) with no volume changes under voltage bias, thereby controlling the spatially averaged conductivity of the film device. A theoretical model to explain the observed hysteretic switching behavior based on electrochemical nonequilibrium thermodynamics is presented, in which the metal-insulator transition results from electrically driven phase separation of Li₄Ti₅O₁₂ and Li₇Ti₅O₁₂. Ability of highly lithiated phase of Li₇Ti₅O₁₂ for Deep Neural Network applications is reported, given the large retentions and symmetry, and opportunity for the low lithiated phase of Li₄Ti₅O₁₂ toward Spiking Neural Network applications, due to the shorter retention and large resistance changes. The findings pave the way for lithium oxides to enable thin-film memristive devices with adjustable symmetry and retention.