Comparison of Echo-Planar Imaging and Compressed Sensing in the Estimation of Flow Metrics from Aortic 4D Flow MR Imaging: A Healthy Volunteer Study.

PURPOSE: Prolonged scanning of time-resolved 3D phase-contrast MRI (4D flow MRI) limits its routine use in clinical practice. An echo-planar imaging (EPI)-based sequence and compressed sensing can reduce the scan duration. We aimed to determine the impact of EPI for 4D flow MRI on the scan duration, image quality, and quantitative flow metrics. METHODS: This was a prospective study of 15 healthy volunteers (all male, mean age 33 ± 5 years). Conventional sensitivity encoding (SENSE), EPI with SENSE (EPI), and compressed SENSE (CS) (reduction factors: 6 and 12, respectively) were scanned.Scan duration, qualitative indexes of image quality, and quantitative flow parameters of net flow volume, maximum flow velocity, wall shear stress (WSS), and energy loss (EL) in the ascending aorta were assessed. Two-dimensional phase-contrast cine MRI (2D-PC) was considered the gold standard of net flow volume and maximum flow velocity. RESULTS: Compared to SENSE, EPI and CS12 shortened scan durations by 71% and 73% (EPI, 4 min 39 sec; CS6, 7 min 29 sec; CS12, 4 min 14 sec; and SENSE, 15 min 51 sec). Visual image quality was significantly better for EPI than for SENSE and CS (P < 0.001). The net flow volumes obtained with SENSE, EPI, and CS12 and those obtained with 2D-PC were correlated well (r = 0.950, 0.871, and 0.850, respectively). However, the maximum velocity obtained with EPI was significantly underestimated (P < 0.010). The average WSS was significantly higher with EPI than with SENSE, CS6, and CS12 (P < 0.001, P = 0.040, and P = 0.012, respectively). The EL was significantly lower with EPI than with CS6 and CS12 (P = 0.002 and P = 0.007, respectively). CONCLUSION: EPI reduced the scan duration, improved visual image quality, and was associated with more accurate net flow volume than CS. However, the flow velocity, WSS, and EL values obtained with EPI and other sequences may not be directly comparable.

Rapid Generation of Murine Bispecific Antibodies Using FAST-IgTM for Preclinical Screening of HER2/CD3 T-Cell Engagers.

Bispecific antibodies (BsAbs) can bind to two different antigens, enabling therapeutic concepts that cannot be achieved with monoclonal antibodies. Immuno-competent mice are essential for validating drug discovery concepts, necessitating the development of surrogate mouse BsAbs. In this study, we explored the potential of FAST-IgTM, a previously reported BsAb technology, for mouse BsAb production. We investigated charge-based orthogonal Fab mutations to facilitate the correct assembly of heavy and light chains of mouse antibodies and employed knobs-into-holes mutations to facilitate the heterodimerization of heavy chains. We combined five anti-CD3 and two anti-HER2 antibodies in mouse IgG1 and IgG2a subclasses. These 20 BsAbs were analyzed using mass spectrometry or ion exchange chromatography to calculate the percentages of BsAbs with correct chain pairing (BsAb yields). Using FAST-Ig, 19 out of the 20 BsAbs demonstrated BsAb yields of 90% or higher after simple protein A purification from transiently expressed antibodies in Expi293F cells. Importantly, the mouse BsAbs maintained their fundamental physicochemical properties and affinity against each antigen. A Jurkat NFAT-luciferase reporter cell assay demonstrated the combined effects of epitope, affinity, and subclasses. Our findings highlight the potential of FAST-Ig technology for efficiently generating mouse BsAbs for preclinical studies.

Efficient production of bispecific antibody by FAST-IgTM and its application to NXT007 for the treatment of hemophilia A.

Efficient production of bispecific antibodies (BsAbs) in single mammalian cells is essential for basic research and industrial manufacturing. However, preventing unwanted pairing of heavy chains (HCs) and light chains (LCs) is a challenging task. To address this, we created an engineering technology for preferential cognate HC/LC and HC/HC paring called FAST-Ig (Four-chain Assembly by electrostatic Steering Technology - Immunoglobulin), and applied it to NXT007, a BsAb for the treatment of hemophilia A. We introduced charged amino-acid substitutions at the HC/LC interface to facilitate the proper assembly for manufacturing a standard IgG-type BsAb. We generated CH1/CL interface-engineered antibody variants that achieved > 95% correct HC/LC pairing efficiency with favorable pharmacological properties and developability. Among these, we selected a design (C3) that allowed us to separate the mis-paired species with an unintended pharmacological profile using ion-exchange chromatography. Crystal structure analysis demonstrated that the C3 design did not affect the overall structure of both Fabs. To determine the final design for HCs-heterodimerization, we compared the stability of charge-based and knobs into hole-based Fc formats in acidic conditions and selected the more stable charge-based format. FAST-Ig was also applicable to stable CHO cell lines for industrial production and demonstrated robust chain pairing with different subclasses of parent BsAbs. Thus, it can be applied to a wide variety of BsAbs both preclinically and clinically.

Variation of the 4 × 1 to 8 × 2 structural transition temperature of quasi-one-dimensional indium chains upon carrier doping from Si(111) substrates.

We investigated the 4 × 1 to 8 × 2 structural transition temperature of quasi-one-dimensional indium chains on the (111) surface of Si substrates possessing various carrier concentrations via low-energy electron diffraction. The transition temperature was found to decrease from 120 K to below 77 K with increasing carrier concentration on both n- and p-type Si(111) substrates. This decrease in the transition temperature was found to be proportional to the shift of the Fermi level, which was numerically evaluated using a one-dimensional charge transfer model of the interface. The obtained results demonstrate that doping of the surface state with both electrons and holes can be readily controlled by judicious selection of Si substrates with appropriate carrier type and concentration.

Activation and splitting of carbon dioxide on the surface of an inorganic electride material.

Activation of carbon dioxide is the most important step in its conversion into valuable chemicals. Surfaces of stable oxide with a low work function may be promising for this purpose. Here we report that the surfaces of the inorganic electride [Ca24Al28O64](4+)(e(-))4 activate and split carbon dioxide at room temperature. This behaviour is attributed to a high concentration of localized electrons in the near-surface region and a corrugation of the surface that can trap oxygen atoms and strained carbon monoxide and carbon dioxide molecules. The [Ca24Al28O64](4+)(e(-))4 surface exposed to carbon dioxide is studied using temperature-programmed desorption, and spectroscopic methods. The results of these measurements, corroborated with ab initio simulations, show that both carbon monoxide and carbon dioxide adsorb on the [Ca24Al28O64](4+)(e(-))4 surface at RT and above and adopt unusual configurations that result in desorption of molecular carbon monoxide and atomic oxygen upon heating.

Quantum interference of Rashba-type spin-split surface state electrons.

We studied the quantum interference of electrons in the Bi (p(x), p(y)) orbital-derived j = 1/2 spin-split surface states at Bi/Ag(111)√3 × âˆš3 surfaces of 10 monolayer thick Ag(111) films on Si(111) substrates. Surface electron standing waves were observed clearly at the energy (E) below the intersection of the two spin-split downward dispersing parabola bands (E(x)). The E dependence of the standing wave pattern reveals the dispersion as the average of the two spin-split surface bands due to the interference between |(k + Δ), ↑> and |-(k - Δ), ↑> [or (|(k - Δ), ↓>) and |-(k + Δ), ↓>] states. In contrast, it was impossible to deduce the dispersion from the standing wave pattern at E ≥ E(x) because the surface electron cannot find its backscattered state with the same spin polarization.

Surface of room-temperature-stable electride [Ca24Al28O64]4+(e-)4: preparation and its characterization by atomic-resolution scanning tunneling microscopy.

The nanocage compound crystal [Ca24Al28O64]4+(e-)4 (C12A7:e-) is a room-temperature-stable electride. Although bulk C12A7:e- exhibits metallic conduction, the surface of an as-prepared sample or one prepared by mechanical fracture in ultrahigh vacuum is almost insulating and exhibits distinct non-ohmic contact. We studied whether the intrinsic surface of this electride exhibits metallic conduction or not by examining various conditions for preparing the intrinsic surface. A combination of sputtering with thermal annealing led to the emergence of metallic conductivity in a specific condition. Suitably prepared surfaces revealed ohmic contact even in an ambient atmosphere. Atomic-resolution scanning tunneling microscopy (STM) images of the surfaces were consistent with a structural model in which the cage structure in the bulk C12A7:e- electride is conserved at the surface.

Dislocation-induced local modulation of the surface states of Ag(111) thin films on Si(111) 7 x 7 substrates.

Local modulation of the Shockley-type surface state was studied around threading dislocations at the surfaces of ultrathin Ag(111) epitaxial films on Si(111) substrates. Scanning tunneling microscope (STM) observations indicated that the wavelength of the surface state electron was shortened around the dislocations in the electron standing wave pattern. Scanning tunneling spectroscopy (STS) revealed that the bottom of the local surface state (E0) shifts downward around the dislocation. The shift in E0 and the lattice displacement Delta u(z) have a linear relation, which indicates that the shift of the surface state is caused by local relaxation of the misfit strain around the dislocation.

Cost of oviposition site selection in a water strider Aquarius paludum insularis: egg mortality increases with oviposition depth.

Females generally avoid selecting sites for oviposition which have a high predation risk to increase offspring survival. Previous studies have focused on costs to ovipositing females. However, although offspring may also incur costs by being oviposited at low predation risk sites, no studies have focused on costs to offspring. Such costs to offspring were examined by using Aquarius paludum insularis, females of which avoid eggs parasitism by ovipositing at deep sites. Deep sites are safe from egg parasitism but may be unsuitable for hatching due to environmental factors. We examined the costs to offspring at deep sites by comparing the hatching rate, the duration to hatching and the proportion of drowned larvae between eggs that were set at three levels of water depth (0 cm, 25 cm and 50 cm depth). While the hatching rate at 50 cm was lower than that at 0 cm, the rate at 25 cm did not differ from that at 0 cm. Duration to hatching and the proportion of drowned larvae did not differ between the three depths. It is suggested that the declining survival rate of A. paludum eggs was due to increased water pressure at greater depth. Such a cost may exist in other species and such an observation may aid in understanding oviposition site selection.