A SPLUNC1 Peptidomimetic Inhibits Orai1 and Reduces Inflammation in a Murine Allergic Asthma Model.

Orai1 is a plasma membrane Ca2+ channel that mediates store-operated Ca2+ entry (SOCE) and regulates inflammation. Short palate lung and nasal epithelial clone 1 (SPLUNC1) is an asthma gene modifier that inhibits Orai1 and SOCE via its C-terminal α6 region. SPLUNC1 levels are diminished in asthma patient airways. Thus, we hypothesized that inhaled α6 peptidomimetics could inhibit Orai1 and reduce airway inflammation in a murine asthma model. To evaluate α6-Orai1 interactions, we used fluorescent assays to measure Ca2+ signaling, Förster resonance energy transfer, fluorescent recovery after photobleaching, immunostaining, total internal reflection microscopy, and Western blotting. To test whether α6 peptidomimetics inhibited SOCE and decreased inflammation in vivo, wild-type and SPLUNC1-/- mice were exposed to house dust mite (HDM) extract with or without α6 peptide. We also performed nebulization, jet milling, and scanning electron microscopy to evaluate α6 for inhalation. SPLUNC1-/- mice had an exaggerated response to HDM. In BAL-derived immune cells, Orai1 levels increased after HDM exposure in SPLUNC1-/- but not wild-type mice. Inhaled α6 reduced Orai1 levels in mice regardless of genotype. In HDM-exposed mice, α6 dose-dependently reduced eosinophilia and neutrophilia. In vitro, α6 inhibited SOCE in multiple immune cell types, and α6 could be nebulized or jet milled without loss of function. These data suggest that α6 peptidomimetics may be a novel, effective antiinflammatory therapy for patients with asthma.

Pubertal Testosterone and Brain Response to Faces in Young Adulthood: An Interplay between Organizational and Activational Effects in Young Men.

According to the organizational-activational hypothesis, the organizational effects of testosterone during (prenatal) brain development moderate the activational effects of adult testosterone on behavior. Accumulating evidence supports the notion that adolescence is another period during which sex hormones organize the nervous system. Here we investigate how pubertal sex hormones moderate the activational effects of adult sex hormones on social cognition in humans. To do so, we recruited a sample of young men (n = 507; age, ∼19 years) from a longitudinal birth cohort and investigated whether testosterone exposure during adolescence (from 9 to 17 years of age) moderates the relation between current testosterone and brain response to faces in young adulthood, as assessed with functional magnetic resonance imaging (fMRI). Our results showed that the cumulative exposure to testosterone during adolescence moderated the relation between adult testosterone and both the mean fMRI response and functional connectivity (i.e., node strength). Specifically, in participants with low exposure to testosterone during puberty, we observed a positive relationship between current testosterone and the brain response to faces; this was not the case for participants with medium and high pubertal testosterone. Furthermore, we observed a stronger relationship between the brain response and current testosterone in parts of the angry-face network associated with (vs without) motion in the eye region of an observed (angry) face. We speculate that pubertal testosterone modulates the relationship between current testosterone and brain response to social cues carried by the eyes and signaling a potential threat.SIGNIFICANCE STATEMENT Accumulating evidence supports the organizational effects of pubertal testosterone, but the body of literature examining these effects on social cognition in humans is in its infancy. With a sample of young men from a longitudinal birth cohort, we showed that the cumulative exposure to testosterone during adolescence moderated the relation between adult testosterone and both the mean BOLD signal change and functional connectivity. Specifically, we observed a positive relationship between adult testosterone and the brain response to faces in participants with low exposure to testosterone during puberty, but not in participants with medium and high pubertal testosterone. Results of further analysis suggest that sensitivity to cues carried by the eyes might underlie the relationship between testosterone and brain response to faces, especially in the context of a potential threat.

Global and Regional Development of the Human Cerebral Cortex: Molecular Architecture and Occupational Aptitudes.

We have carried out meta-analyses of genome-wide association studies (GWAS) (n = 23 784) of the first two principal components (PCs) that group together cortical regions with shared variance in their surface area. PC1 (global) captured variations of most regions, whereas PC2 (visual) was specific to the primary and secondary visual cortices. We identified a total of 18 (PC1) and 17 (PC2) independent loci, which were replicated in another 25 746 individuals. The loci of the global PC1 included those associated previously with intracranial volume and/or general cognitive function, such as MAPT and IGF2BP1. The loci of the visual PC2 included DAAM1, a key player in the planar-cell-polarity pathway. We then tested associations with occupational aptitudes and, as predicted, found that the global PC1 was associated with General Learning Ability, and the visual PC2 was associated with the Form Perception aptitude. These results suggest that interindividual variations in global and regional development of the human cerebral cortex (and its molecular architecture) cascade-albeit in a very limited manner-to behaviors as complex as the choice of one's occupation.

Hyperspectral multiphoton microscopy for in vivo visualization of multiple, spectrally overlapped fluorescent labels.

The insensitivity of multiphoton microscopy to optical scattering enables high-resolution, high-contrast imaging deep into tissue, including in live animals. Scattering does, however, severely limit the use of spectral dispersion techniques to improve spectral resolution. In practice, this limited spectral resolution together with the need for multiple excitation wavelengths to excite different fluorophores limits multiphoton microscopy to imaging a few, spectrally-distinct fluorescent labels at a time, restricting the complexity of biological processes that can be studied. Here, we demonstrate a hyperspectral multiphoton microscope that utilizes three different wavelength excitation sources together with multiplexed fluorescence emission detection using angle-tuned bandpass filters. This microscope maintains scattering insensitivity, while providing high enough spectral resolution on the emitted fluorescence and capitalizing on the wavelength-dependent nonlinear excitation of fluorescent dyes to enable clean separation of multiple, spectrally overlapping labels, in vivo. We demonstrated the utility of this instrument for spectral separation of closely-overlapped fluorophores in samples containing ten different colors of fluorescent beads, live cells expressing up to seven different fluorescent protein fusion constructs, and in multiple in vivo preparations in mouse cortex and inflamed skin with up to eight different cell types or tissue structures distinguished.

Blind deconvolution in model-based iterative reconstruction for CT using a normalized sparsity measure.

Model-based iterative reconstruction techniques for CT that include a description of the noise statistics and a physical forward model of the image formation process have proven to increase image quality for many applications. Specifically, including models of the system blur into the physical forward model and thus implicitly performing a deconvolution of the projections during tomographic reconstruction, could demonstrate distinct improvements, especially in terms of resolution. However, the results strongly rely on an exact characterization of all components contributing to the system blur. Such characterizations can be laborious and even a slight mismatch can diminish image quality significantly. Therefore, we introduce a novel objective function, which enables us to jointly estimate system blur parameters during tomographic reconstruction. Conventional objective functions are biased in terms of blur and can yield lowest cost to blurred reconstructions with low noise levels. A key feature of our objective function is a new normalized sparsity measure for CT based on total-variation regularization, constructed to be less biased in terms of blur. We outline a solving strategy for jointly recovering low-dimensional blur parameters during tomographic reconstruction. We perform an extensive simulation study, evaluating the performance of the regularization and the dependency of the different parts of the objective function on the blur parameters. Scenarios with different regularization strengths and system blurs are investigated, demonstrating that we can recover the blur parameter used for the simulations. The proposed strategy is validated and the dependency of the objective function with the number of iterations is analyzed. Finally, our approach is experimentally validated on test-bench data of a human wrist phantom, where the estimated blur parameter coincides well with visual inspection. Our findings are not restricted to attenuation-based CT and may facilitate the recovery of more complex imaging model parameters.

Model-based material decomposition with a penalized nonlinear least-squares CT reconstruction algorithm.

Spectral information in CT may be used for material decomposition to produce accurate reconstructions of material density and to separate materials with similar overall attenuation. Traditional methods separate the reconstruction and decomposition steps, often resulting in undesirable trade-offs (e.g. sampling constraints, a simplified spectral model). In this work, we present a model-based material decomposition algorithm which performs the reconstruction and decomposition simultaneously using a multienergy forward model. In a kV-switching simulation study, the presented method is capable of reconstructing iodine at 0.5 mg ml-1 with a contrast-to-noise ratio greater than two, as compared to 3.0 mg ml-1 for image domain decomposition. The presented method also enables novel acquisition methods, which was demonstrated in this work with a combined kV-switching/split-filter acquisition explored in simulation and physical test bench studies. This novel design used four spectral channels to decompose three materials: water, iodine, and gadolinium. In simulation, the presented method accurately reconstructed concentration value estimates with RMSE values of 4.86 mg ml-1 for water, 0.108 mg ml-1 for iodine and 0.170 mg ml-1 for gadolinium. In test-bench data, the RMSE values were 134 mg ml-1, 5.26 mg ml-1 and 1.85 mg ml-1, respectively. These studies demonstrate the ability of model-based material decomposition to produce accurate concentration estimates in challenging spatial/spectral sampling acquisitions.

Local response prediction in model-based CT material decomposition.

Spectral CT is an emerging modality that permits material decomposition and density estimation through the use of energy-dependent information in measurements. Direct model-based material decomposition algorithms have been developed that incorporate statistical models and advanced regularization schemes to improve density estimates and lower exposure requirements. However, understanding and control of the relationship between regularization and image properties is complex with interactions between spectral channels and material bases. In particular, regularization in one material basis can affect the image properties of other material bases, and vice versa. In this work, we derived a closed-form set of local impulse responses for the solutions to a general, regularized, model-based material decomposition (MBMD) objective. These predictors quantify both the spatial resolution in each material image as well as the influence of regularization of one material basis on other material images. This information can be used prospectively to tune regularization parameters for specific imaging goals.

Optimized Spatial-Spectral CT for Multi-Material Decomposition.

Spectral CT is an emerging modality that uses a data acquisition scheme with varied spectral responses to provide enhanced material discrimination in addition to the structural information of conventional CT. Existing clinical and preclinical designs with this capability include kV-switching, split-filtration, and dual-layer detector systems to provide two spectral channels of projection data. In this work, we examine an alternate design based on a spatial-spectral filter. This source-side filter is made up a linear array of materials that divide the incident x-ray beam into spectrally varied beamlets. This design allows for any number of spectral channels; however, each individual channel is sparse in the projection domain. Model-based iterative reconstruction methods can accommodate such sparse spatial-spectral sampling patterns and allow for the incorporation of advanced regularization. With the goal of an optimized physical design, we characterize the effects of design parameters including filter tile order and filter tile width and their impact on material decomposition performance. We present results of numerical simulations that characterize the impact of each design parameter using a realistic CT geometry and noise model to demonstrate feasibility. Results for filter tile order show little change indicating that filter order is a low-priority design consideration. We observe improved performance for narrower filter widths; however, the performance drop-off is relatively flat indicating that wider filter widths are also feasible designs.

Model-based Multi-material Decomposition using Spatial-Spectral CT Filters.

Spectral CT with multiple contrast agents has been enabled by energy-discriminating detectors with multiple spectral channels. We propose a new approach that uses spatial-spectral filters to provide multiple beamlets with different incident spectra for spectral channels based on "source-side" control. Since these spatial-spectral filters yield spectral channels that are sparse, we adopt model-based material decomposition to directly reconstruct material densities from projection data. Simulation studies in three-and four-material decomposition experiments show the underlying feasibility of the spatial-spectral filtering technique. This methodology has the potential to facilitate imaging of multiple contrast agents simultaneously with relatively simple hardware, or to improve spectral CT performance via combination with other established spectral CT methods for additional control and flexibility.

High-Fidelity Modeling of Detector Lag and Gantry Motion in CT Reconstruction.

Detector lag and gantry motion during x-ray exposure and integration both result in azimuthal blurring in CT reconstructions. These effects can degrade image quality both for high-resolution features as well as low-contrast details. In this work we consider a forward model for model-based iterative reconstruction (MBIR) that is sufficiently general to accommodate both of these physical effects. We integrate this forward model in a penalized, weighted, nonlinear least-square style objective function for joint reconstruction and correction of these blur effects. We show that modeling detector lag can reduce/remove the characteristic lag artifacts in head imaging in both a simulation study and physical experiments. Similarly, we show that azimuthal blur ordinarily introduced by gantry motion can be mitigated with proper reconstruction models. In particular, we find the largest image quality improvement at the periphery of the field-of-view where gantry motion artifacts are most pronounced. These experiments illustrate the generality of the underlying forward model, suggesting the potential application in modeling a number of physical effects that are traditionally ignored or mitigated through pre-corrections to measurement data.

A General CT Reconstruction Algorithm for Model-Based Material Decomposition.

Material decomposition in CT has the potential to reduce artifacts and improve quantitative accuracy by utilizing spectral models and multi-energy scans. In this work we present a novel Model-Based Material Decomposition (MBMD) method based on an existing iterative reconstruction algorithm derived from a general non-linear forward model. A digital water phantom with inserts containing different concentrations of calcium was scanned on a kV switching system. We used the presented method to simultaneously reconstruct water and calcium material density images, and compared the results to an image domain and a projection domain decomposition method. When switching voltage every other frame, MBMD resulted in more accurate water and calcium concentration values than the image domain decomposition method, and was just as accurate as the projection domain decomposition method. In a second, slower, kV switching scheme (changing voltage every ten frames) which precluded the use of traditional projection domain based methods, MBMD continued to produce quantitatively accurate reconstructions. Finally, we present a preliminary study applying MBMD to a water phantom containing vials of different concentrations of K2HPO4 which was scanned on a cone-beam CT test bench. Both the fast and slow (emulated) kV switching schemes resulted in similar reconstructions, indicating MBMD's robustness to challenging acquisition schemes. Additionally, the K2HPO4 concentration ratios between the vials were accurately represented in the reconstructed K2HPO4 density image.

Penalized-Likelihood Reconstruction With High-Fidelity Measurement Models for High-Resolution Cone-Beam Imaging.

We present a novel reconstruction algorithm based on a general cone-beam CT forward model, which is capable of incorporating the blur and noise correlations that are exhibited in flat-panel CBCT measurement data. Specifically, the proposed model may include scintillator blur, focal-spot blur, and noise correlations due to light spread in the scintillator. The proposed algorithm (GPL-BC) uses a Gaussian Penalized-Likelihood objective function, which incorporates models of blur and correlated noise. In a simulation study, GPL-BC was able to achieve lower bias as compared with deblurring followed by FDK as well as a model-based reconstruction method without integration of measurement blur. In the same study, GPL-BC was able to achieve better line-pair reconstructions (in terms of segmented-image accuracy) as compared with deblurring followed by FDK, a model-based method without blur, and a model-based method with blur but not noise correlations. A prototype extremities quantitative cone-beam CT test-bench was used to image a physical sample of human trabecular bone. These data were used to compare reconstructions using the proposed method and model-based methods without blur and/or correlation to a registered CT image of the same bone sample. The GPL-BC reconstructions resulted in more accurate trabecular bone segmentation. Multiple trabecular bone metrics, including trabecular thickness (Tb.Th.) were computed for each reconstruction approach as well as the CT volume. The GPL-BC reconstruction provided the most accurate Tb.Th. measurement, 0.255 mm, as compared with the CT derived value of 0.193 mm, followed by the GPL-B reconstruction, the GPL-I reconstruction, and then the FDK reconstruction (0.271 mm, 0.309 mm, and 0.335 mm, respectively).

Nonlinear Statistical Reconstruction for Flat-Panel Cone-Beam CT with Blur and Correlated Noise Models.

Flat-panel cone-beam CT (FP-CBCT) is a promising imaging modality, partly due to its potential for high spatial resolution reconstructions in relatively compact scanners. Despite this potential, FP-CBCT can face difficulty resolving important fine scale structures (e.g, trabecular details in dedicated extremities scanners and microcalcifications in dedicated CBCT mammography). Model-based methods offer one opportunity to improve high-resolution performance without any hardware changes. Previous work, based on a linearized forward model, demonstrated improved performance when both system blur and spatial correlations characteristics of FP-CBCT systems are modeled. Unfortunately, the linearized model relies on a staged processing approach that complicates tuning parameter selection and can limit the finest achievable spatial resolution. In this work, we present an alternative scheme that leverages a full nonlinear forward model with both system blur and spatially correlated noise. A likelihood-based objective function is derived from this forward model and we derive an iterative optimization algorithm for its solution. The proposed approach is evaluated in simulation studies using a digital extremities phantom and resolution-noise trade-offs are quantitatively evaluated. The correlated nonlinear model outperformed both the uncorrelated nonlinear model and the staged linearized technique with up to a 86% reduction in variance at matched spatial resolution. Additionally, the nonlinear models could achieve finer spatial resolution (correlated: 0.10 mm, uncorrelated: 0.11 mm) than the linear correlated model (0.15 mm), and traditional FDK (0.40 mm). This suggests the proposed nonlinear approach may be an important tool in improving performance for high-resolution clinical applications.

Design of dual multiple aperture devices for dynamical fluence field modulated CT.

A Multiple Aperture Device (MAD) is a novel x-ray beam modulator that uses binary filtration on a fine scale to spatially modulate an x-ray beam. Using two MADs in series enables a large variety of fluence profiles by shifting the MADS relative to each other. This work details the design and control of dual MADs for a specific class of desired fluence patterns. Specifically, models of MAD operation are integrated into a best fit objective followed by CMA-ES optimization. To illustrate this framework we demonstrate the design process for an abdominal phantom with the goal of uniform detected signal. Achievable fluence profiles show good agreement with target fluence profiles, and the ability to flatten projections when a phantom is scanned is demonstrated. Simulated data reconstruction using traditional tube current modulation (TCM) and MAD filtering with TCM are investigated with the dual MAD system demonstrating more uniformity in noise and illustrating the potential for dose reduction under a maximum noise level constraint.

Modeling Shift-Variant X-Ray Focal Spot Blur for High-Resolution Flat-Panel Cone-Beam CT.

Flat-panel cone-beam CT (CBCT) has been applied clinically in a number of high-resolution applications. Increasing geometric magnification can potentially improve resolution, but also increases blur due to an extended x-ray focal-spot. We present a shift-variant focal-spot blur model and incorporate it into a model-based iterative-reconstruction algorithm. We apply this algorithm to simulation and CBCT test-bench data. In a trabecular bone simulation study, we find traditional reconstruction approaches without a blur model exhibit shift-variant resolution properties that depend greatly on the acquisition protocol (e.g. short vs. full scans) and the anode angles of the rays used to reconstruct a particular region. For physical CBCT experiments focal spot blur was characterized and a spatial resolution phantom was scanned and reconstructed. In both experiments image quality using the shift-variant model was significantly improved over approaches that modeled no blur or only a shift-invariant blur, suggesting a potential means to overcome traditional CBCT spatial resolution and system design limitations.

Model-based iterative reconstruction for flat-panel cone-beam CT with focal spot blur, detector blur, and correlated noise.

While model-based reconstruction methods have been successfully applied to flat-panel cone-beam CT (FP-CBCT) systems, typical implementations ignore both spatial correlations in the projection data as well as system blurs due to the detector and focal spot in the x-ray source. In this work, we develop a forward model for flat-panel-based systems that includes blur and noise correlation associated with finite focal spot size and an indirect detector (e.g. scintillator). This forward model is used to develop a staged reconstruction framework where projection data are deconvolved and log-transformed, followed by a generalized least-squares reconstruction that utilizes a non-diagonal statistical weighting to account for the correlation that arises from the acquisition and data processing chain. We investigate the performance of this novel reconstruction approach in both simulated data and in CBCT test-bench data. In comparison to traditional filtered backprojection and model-based methods that ignore noise correlation, the proposed approach yields a superior noise-resolution tradeoff. For example, for a system with 0.34 mm FWHM scintillator blur and 0.70 FWHM focal spot blur, using the correlated noise model instead of an uncorrelated noise model increased resolution by 42% (with variance matched at 6.9 × 10(-8) mm(-2)). While this advantage holds across a wide range of systems with differing blur characteristics, the improvements are greatest for systems where source blur is larger than detector blur.

Integration of Component Knowledge in Penalized-Likelihood Reconstruction with Morphological and Spectral Uncertainties.

Previous investigations [1-3] have demonstrated that integrating specific knowledge of the structure and composition of components like surgical implants, devices, and tools into a model-based reconstruction framework can improve image quality and allow for potential exposure reductions in CT. Using device knowledge in practice is complicated by uncertainties in the exact shape of components and their particular material composition. Such unknowns in the morphology and attenuation properties lead to errors in the forward model that limit the utility of component integration. In this work, a methodology is presented to accommodate both uncertainties in shape as well as unknown energy-dependent attenuation properties of the surgical devices. This work leverages the so-called known-component reconstruction (KCR) framework [1] with a generalized deformable registration operator and modifications to accommodate a spectral transfer function in the component model. Moreover, since this framework decomposes the object into separate background anatomy and "known" component factors, a mixed fidelity forward model can be adopted so that measurements associated with projections through the surgical devices can be modeled with much greater accuracy. A deformable KCR (dKCR) approach using the mixed fidelity model is introduced and applied to a flexible wire component with unknown structure and composition. Image quality advantages of dKCR over traditional reconstruction methods are illustrated in cone-beam CT (CBCT) data acquired on a testbench emulating a 3D-guided needle biopsy procedure - i.e., a deformable component (needle) with strong energy-dependent attenuation characteristics (steel) within a complex soft-tissue background.

Generalized Least-Squares CT Reconstruction with Detector Blur and Correlated Noise Models.

The success and improved dose utilization of statistical reconstruction methods arises, in part, from their ability to incorporate sophisticated models of the physics of the measurement process and noise. Despite the great promise of statistical methods, typical measurement models ignore blurring effects, and nearly all current approaches make the presumption of independent measurements - disregarding noise correlations and a potential avenue for improved image quality. In some imaging systems, such as flat-panel-based cone-beam CT, such correlations and blurs can be a dominant factor in limiting the maximum achievable spatial resolution and noise performance. In this work, we propose a novel regularized generalized least-squares reconstruction method that includes models for both system blur and correlated noise in the projection data. We demonstrate, in simulation studies, that this approach can break through the traditional spatial resolution limits of methods that do not model these physical effects. Moreover, in comparison to other approaches that attempt deblurring without a correlation model, superior noise-resolution trade-offs can be found with the proposed approach.

Iterative CT Reconstruction using Models of Source and Detector Blur and Correlated Noise.

Statistical model-based reconstruction methods derive much of their advantage over traditional methods through more accurate forward models of the imaging system. Typical forward models fail to integrate two important aspects of real imaging systems: system blur and noise correlations in the measurements. This work develops an approach that models both aspects using a two-stage approach that includes a regularization deblurring operation followed by generalized penalized weighted least-squares reconstruction. Different reconstruction noise models including standard uncorrelated and correlated presumptions were explored. Moreover, different imaging systems were investigated in which blur was dominated by source effects, dominated by detector effects, or by a combination of source and detector blur. The proposed reconstruction approach that models the correlated noise demonstrated the best performance across all scenarios with the greatest benefits for increased source blur and for reconstructions with finer spatial resolution. This suggests potential application of the method for high resolution systems like dedicated flat-panel cone-beam CT (e.g., head, extremity, dental, mammography scanners) where system resolution is limited by both source and detector blur effects and noise correlations in measurement data are traditionally ignored.