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Ultrasound (US) imaging is used to assess cardiac disease by assessing the geometry and function of the heart utilizing its high spatial and temporal resolution. However, because of physical constraints, drawbacks of US include limited field-of-view, refraction, resolution and contrast anisotropy. These issues cannot be resolved when using a single probe. Here, an interleaved multi-perspective 2-D US imaging system was introduced, aiming at improved imaging of the left ventricle (LV) of the heart by acquiring US data from two separate phased array probes simultaneously at a high frame rate. In an ex-vivo experiment of a beating porcine heart, parasternal long-axis and apical views of the left ventricle were acquired using two phased array probes. Interleaved multi-probe US data were acquired at a frame rate of 170 frames per second (FPS) using diverging wave imaging under 11 angles. Image registration and fusion algorithms were developed to align and fuse the US images from two different probes. First- and second-order speckle statistics were computed to characterize the resulting probability distribution function and point spread function of the multi-probe image data. First-order speckle analysis showed less overlap of the histograms (reduction of 34.4%) and higher contrast-to-noise ratio (CNR, increase of 27.3%) between endocardium and myocardium in the fused images. Autocorrelation results showed an improved and more isotropic resolution for the multi-perspective images (single-perspective: 0.59 mm × 0.21 mm, multi-perspective: 0.35 mm × 0.18 mm). Moreover, mean gradient (MG) (increase of 74.4%) and entropy (increase of 23.1%) results indicated that image details of the myocardial tissue can be better observed after fusion. To conclude, interleaved multi-perspective high frame rate US imaging was developed and demonstrated in an ex-vivo experimental setup, revealing enlarged field-of-view, and improved image contrast and resolution of cardiac images.

Fc galactosylation is a critical quality attribute for anti-tumor recombinant immunoglobulin G (IgG)-based monoclonal antibody (mAb) therapeutics with complement-dependent cytotoxicity (CDC) as the mechanism of action. Although the correlation between galactosylation and CDC has been known, the underlying structure–function relationship is unclear. Heterogeneity of the Fc N-glycosylation produced by Chinese hamster ovary (CHO) cell culture biomanufacturing process leads to variable CDC potency. Here, we derived a kinetic model of galactose transfer reaction in the Golgi apparatus and used this model to determine the correlation between differently galactosylated species from CHO cell culture process. The model was validated by a retrospective data analysis of more than 800 historical samples from small-scale and large-scale CHO cell cultures. Furthermore, using various analytical technologies, we discovered the molecular basis for Fc glycan terminal galactosylation changing the three-dimensional conformation of the Fc, which facilitates the IgG1 hexamerization, thus enhancing C1q avidity and subsequent complement activation. Our study offers insight into the formation of galactosylated species, as well as a novel three-dimensional understanding of the structure–function relationship of terminal galactose to complement activation in mAb therapeutics.

Cadherins are calcium-dependent, transmembrane adhesion molecules that assemble through direct noncovalent association of their N-terminal extracellular modular domains. As the transmembrane component of adherens junctions, they indirectly link adherent cells' actin cytoskeletons. Here, we investigate the most distal extracellular domain of neural cadherin (N-cadherin), a protein required at excitatory synapses, the site of long-term potentiation. This domain is the site of the adhesive interface, and it forms a dimer spontaneously without binding calcium, a surprising finding given that calcium binding is required for proper physiological function. A critical tryptophan at position 2, W2, provides a spectroscopic probe for the "closed" monomer and strand-swapped dimer. Spectroscopic studies show that W2 remains docked in the two forms but has a different apparent interaction with the hydrophobic pocket. Size-exclusion chromatography was used to measure the levels of the monomer and dimer over time to study the kinetics and equilibria of the unexpected spontaneous dimer formation ( K

1-Butyl-3-methylimidazolium hydrogen sulfate ([Bmim][HSO4]) was employed to catalyze the in-situ transesterification of wet Nannochloropsis oceanica. Algal cell wall was dissolved by [Bmim][HSO4] according to TEM analysis of untreated Nannochloropsis sp. cells and lipid extracted algae (LEA). The temperature and time are favorable to biodiesel production in 100–200 °C, time of 0–70 min, with methanol: algae, however, it decreased to 10.64% at time of 90 min. The biodiesel and energy production of [Bmim][HSO4] catalyzed in-situ transesterification varied due to the competition of catalytic property and algal dissolution ability with variation of water content. The catalytic property of [Bmim][HSO4] was the dominate parameter affecting biodiesel production, therefore, total energy production of in-situ transesterification, as a parameter of biodiesel production, was slightly increased from 13.12 kJ to 14.51KJ with water content of wet algae varied from 0 to 15 wt%. However, it decreased to 8.37 at water content of 20 wt% due to the weakened algal dissolution ability, which was proved by the decrease of hydrogen bond acceptor capacity(β) from 0.942858 to 0.942851 in water content of 0–30 wt%. The decomposition of carbohydrates and proteins in IL – water mixtures enhanced biodiesel production according to elemental analysis of LEA.

The non-contact torque sensing application is used widely in modern measurement, but now the structure of torque measurement system was found to be relatively complicated. In this work, a non-contact torque measurement method based on the equivalent theory of the magnetic field and stress is proposed to detect the rotation angle of magnetization using thermodynamic equilibrium principle, and a theoretical model of equivalent stress and rotation angle of magnetization is established. The sensitivity of the torque measurement system reached 28.4 mV/(Nm) and the maximum nonlinearity error is 1.3%FS through the experiment, which verifies the feasibility of the torque measurement method.

Human glucokinase (GCK) acts as the body's primary glucose sensor and plays a critical role in glucose homeostatic maintenance. Gain-of-function mutations in gck produce hyperactive enzyme variants that cause congenital hyperinsulinism. Prior biochemical and biophysical studies suggest that activated disease variants can be segregated into two mechanistically distinct classes, termed α-type and β-type. Steady-state viscosity variation studies indicate that the kcat values of wild-type GCK and an α-type variant are partially diffusion-limited, whereas the kcat value of a β-type variant is viscosity-independent. Transient-state chemical quench-flow analyses demonstrate that wild-type GCK and the α-type variant display burst kinetics, whereas the β-type variant lacks a burst phase. Comparative hydrogen-deuterium exchange mass spectrometry of unliganded enzymes demonstrates that a disordered active site loop, which folds upon binding of glucose, is protected from exchange in the α-type variant. The α-type variant also displays an increased level of exchange within a β-strand located near the enzyme's hinge region, which becomes more solvent-exposed upon glucose binding. In contrast, β-type activation causes no substantial difference in global or local exchange relative to that of unliganded, wild-type GCK. Together, these results demonstrate that α-type activation results from a shift in the conformational ensemble of unliganded GCK toward a state resembling the glucose-bound conformation, whereas β-type activation is attributable to an accelerated rate of product release. This work elucidates the molecular basis of naturally occurring, activated GCK disease variants and provides insight into the structural and dynamic origins of GCK's unique kinetic cooperativity.

The internal structure of off-axis three-mirror system is commonly complex. The mirror installation error in assembly always affects the imaging line-of-sight and further degrades the image quality. Due to the complexity of the optical path in off-axis three-mirror optical system, the straightforward theoretical analysis on the variations of imaging line-of-sight is extremely difficult. In order to simplify the theoretical analysis, an equivalent single-mirror system is proposed and presented in this paper. In addition, the mathematical model of single-mirror system is established and the accurate expressions of imaging coordinate are derived. Utilizing the simulation software ZEMAX, off-axis three-mirror model and single-mirror model are both established. By adjusting the position of mirror and simulating the line-of-sight rotation of optical system, the variations of imaging coordinates are clearly observed. The final simulation results include: in off-axis three-mirror system, the varying sensitivity of the imaging coordinate to the rotation of line-of-sight is approximately 30 um/″; in single-mirror system, the varying sensitivity of the imaging coordinate to the rotation of line-of-sight is 31.5 um/″. Compared to the simulation results of the off-axis three-mirror model, the 5p relative error of single-mirror model analysis highly satisfies the requirement of equivalent analysis and also verifies its validity. This paper presents a new method to analyze the installation error of the mirror in the off-axis three-mirror system influencing on the imaging line-of-sight. Moreover, the off-axis three-mirror model is totally equivalent to the single-mirror model in theoretical analysis.

Acidosis within tumor and kidney tissues has previously been quantitatively measured using a molecular imaging technique known as acidoCEST MRI. The previous studies used iopromide and iopamidol, two iodinated contrast agents that are approved for clinical CT diagnoses and have been repurposed for acidoCEST MRI studies. We aimed to compare the performance of the two agents for measuring pH by optimizing image acquisition conditions, correlating pH with a ratio of CEST effects from an agent, and evaluating the effects of concentration, endogenous T1 relaxation time and temperature on the pH-CEST ratio correlation for each agent. These results showed that the two agents had similar performance characteristics, although iopromide produced a pH measurement with a higher dynamic range while iopamidol produced a more precise pH measurement. We then compared the performance of the two agents to measure in vivo extracellular pH (pHe) within xenograft tumor models of Raji lymphoma and MCF-7 breast cancer. Our results showed that the pHe values measured with each agent were not significantly different. Also, iopromide consistently measured a greater region of the tumor relative to iopamidol in both tumor models. Therefore, an iodinated contrast agent for acidoCEST MRI should be selected based on the measurement properties needed for a specific biomedical study and the pharmacokinetic properties of a specific tumor model.

In order to improve the accuracy of ultrasonic phased array focusing time delay, analyzing the original interpolation Cascade-Integrator-Comb (CIC) filter, an 8× interpolation CIC filter parallel algorithm was proposed, so that interpolation and multichannel decomposition can simultaneously process. Moreover, we summarized the general formula of arbitrary multiple interpolation CIC filter parallel algorithm and established an ultrasonic phased array focusing time delay system based on 8× interpolation CIC filter parallel algorithm. Improving the algorithmic structure, 12.5% of addition and 29.2% of multiplication was reduced, meanwhile the speed of computation is still very fast. Considering the existing problems of the CIC filter, we compensated the CIC filter; the compensated CIC filter's pass band is flatter, the transition band becomes steep, and the stop band attenuation increases. Finally, we verified the feasibility of this algorithm on Field Programming Gate Array (FPGA). In the case of system clock is 125 MHz, after 8× interpolation filtering and decomposition, time delay accuracy of the defect echo becomes 1 ns. Simulation and experimental results both show that the algorithm we proposed has strong feasibility. Because of the fast calculation, small computational amount and high resolution, this algorithm is especially suitable for applications with high time delay accuracy and fast detection.

Summary The Saccharomyces cerevisiae (Sc) R2TP complex affords an Hsp90-mediated and nucleotide-driven chaperone activity to proteins of small ribonucleoprotein particles (snoRNPs). The current lack of structural information on the ScR2TP complex, however, prevents a mechanistic understanding of this biological process. We characterized the structure of the ScR2TP complex made up of two AAA+ ATPases, Rvb1/2p, and two Hsp90 binding proteins, Tah1p and Pih1p, and its interaction with the snoRNP protein Nop58p by a combination of analytical ultracentrifugation, isothermal titration calorimetry, chemical crosslinking, hydrogen-deuterium exchange, and cryoelectron microscopy methods. We find that Pih1p-Tah1p interacts with Rvb1/2p cooperatively through the nucleotide-sensitive domain of Rvb1/2p. Nop58p further binds Pih1p-Tahp1 on top of the dome-shaped R2TP. Consequently, nucleotide binding releases Pih1p-Tah1p from Rvb1/2p, which offers a mechanism for nucleotide-driven binding and release of snoRNP intermediates.

Acidosis within tumor and kidney tissues has previously been quantitatively measured using a molecular imaging technique known as acidoCEST MRI. The previous studies used iopromide and iopamidol, two iodinated contrast agents that are approved for clinical CT diagnoses and have been repurposed for acidoCEST MRI studies. We aimed to compare the performance of the two agents for measuring pH by optimizing image acquisition conditions, correlating pH with a ratio of CEST effects from an agent, and evaluating the effects of concentration, endogenous T1 relaxation time and temperature on the pH-CEST ratio correlation for each agent. These results showed that the two agents had similar performance characteristics, although iopromide produced a pH measurement with a higher dynamic range while iopamidol produced a more precise pH measurement. We then compared the performance of the two agents to measure in vivo extracellular pH (pHe) within xenograft tumor models of Raji lymphoma and MCF-7 breast cancer. Our results showed that the pHe values measured with each agent were not significantly different. Also, iopromide consistently measured a greater region of the tumor relative to iopamidol in both tumor models. Therefore, an iodinated contrast agent for acidoCEST MRI should be selected based on the measurement properties needed for a specific biomedical study and the pharmacokinetic properties of a specific tumor model.