Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.

Optical tissue clearing and three-dimensional (3D) immunofluorescence (IF) microscopy is transforming imaging of the complex tumor microenvironment (TME). However, current 3D IF microscopy has restricted multiplexity; only 3 or 4 cellular and noncellular TME components can be localized in cleared tumor tissue. Here we report a light-emitting diode (LED) photobleaching method and its application for 3D multiplexed optical mapping of the TME. We built a high-power LED light irradiation device and temperature-controlled chamber for completely bleaching fluorescent signals throughout optically cleared tumor tissues without compromise of tissue and protein antigen integrity. With newly developed tissue mounting and selected region-tracking methods, we established a cyclic workflow involving IF staining, tissue clearing, 3D confocal microscopy, and LED photobleaching. By registering microscope channel images generated through 3 work cycles, we produced 8-plex image data from individual 400 µm-thick tumor macrosections that visualize various vascular, immune, and cancer cells in the same TME at tissue-wide and cellular levels in 3D. Our method was also validated for quantitative 3D spatial analysis of cellular remodeling in the TME after immunotherapy. These results demonstrate that our LED photobleaching system and its workflow offer a novel approach to increase the multiplexing power of 3D IF microscopy for studying tumor heterogeneity and response to therapy.

Cardiovascular magnetic resonance characterization of myocardial tissue injury in a miniature swine model of cancer therapy-related cardiovascular toxicity.

BACKGROUND: Left ventricular ejection fraction (LVEF) is the most commonly clinically used imaging parameter for assessing cancer therapy-related cardiac dysfunction (CTRCD). However, LVEF declines may occur late, after substantial injury. This study sought to investigate cardiovascular magnetic resonance (CMR) imaging markers of subclinical cardiac injury in a miniature swine model. METHODS: Female Yucatan miniature swine (n = 14) received doxorubicin (2 mg/kg) every 3 weeks for 4 cycles. CMR, including cine, tissue characterization via T1 and T2 mapping, and late gadolinium enhancement (LGE) were performed on the same day as doxorubicin administration and 3 weeks after the final chemotherapy cycle. In addition, magnetic resonance spectroscopy (MRS) was performed during the 3 weeks after the final chemotherapy in 7 pigs. A single CMR and MRS exam were also performed in 3 Yucatan miniature swine that were age- and weight-matched to the final imaging exam of the doxorubicin-treated swine to serve as controls. CTRCD was defined as histological early morphologic changes, including cytoplasmic vacuolization and myofibrillar loss of myocytes, based on post-mortem analysis of humanely euthanized pigs after the final CMR exam. RESULTS: Of 13 swine completing 5 serial CMR scans, 10 (77%) had histological evidence of CTRCD. Three animals had neither histological evidence nor changes in LVEF from baseline. No absolute LVEF <40% or LGE was observed. Native T1, extracellular volume (ECV), and T2 at 12 weeks were significantly higher in swine with CTRCD than those without CTRCD (1178 ms vs. 1134 ms, p = 0.002, 27.4% vs. 24.5%, p = 0.03, and 38.1 ms vs. 36.4 ms, p = 0.02, respectively). There were no significant changes in strain parameters. The temporal trajectories in native T1, ECV, and T2 in swine with CTRCD showed similar and statistically significant increases. At the same time, there were no differences in their temporal changes between those with and without CTRCD. MRS myocardial triglyceride content substantially differed among controls, swine with and without CTRCD (0.89%, 0.30%, 0.54%, respectively, analysis of variance, p = 0.01), and associated with the severity of histological findings and incidence of vacuolated cardiomyocytes. CONCLUSION: Serial CMR imaging alone has a limited ability to detect histologic CTRCD beyond LVEF. Integrating MRS myocardial triglyceride content may be useful for detection of early potential CTRCD.

Comparison of 14 respiratory pathogens among hospitalized children during and after the COVID-19 outbreak in Chaoshan area.

BACKGROUND: Since January 2020, measures has been adopted in the Chaoshan area to limit the spread of COVID-19. Restrictions were removed after August 2020. At the same time, children returned to school. We previously reported the changes of 14 main respiratory pathogens in hospitalized children before and during the COVID-19 outbreak in Chaoshan area. However, the changes of respiratory pathogen spectrum in hospitalized children after the epidemic are still unknown, which will be elucidated in this study. METHODS: There are 6201 children hospitalized with respiratory tract infection were enrolled in the study, which were divided into two groups: 2533 from outbreak group (1 January 2020-31 December 2020), and 3668 from post-outbreak group (1 January 2021-31 December 2021). Pharyngeal swab samples were collected. 14 respiratory tract pathogens were detected by liquid chip technology. RESULTS: The positive rate of pathogen detection is significantly lower in the outbreak group (65.42%, 1657/2533) than that in the post-outbreak group (70.39%, 2582/3668; χ2 = 17.15, P < 0.05). The Influenza A virus (FluA) detection rate was 1.9% (49) in 2020, but 0% (0) in 2021. The detection rates of Bordetella pertussis (BP) decreased from 1.4% (35) in 2020 to 0.5% (17) in 2021. In contrast, the detection rates of  Influenza B virus (FluB), Cytomegalovirus (CMV), Haemophilus influenzae (HI), Streptococcus pneumoniae (SP) increased from 0.3% (8), 24.7% (626), 2.0% (50) and 19.4% (491) in 2020 to 3.3% (121), 27.9% (1025), 4.6% (169), 22.8% (836) in 2021, respectively (P < 0.01). CONCLUSIONS: The detection rates of pathogens such as FluA, FluB, CMV, HI, SP, BP were statistically different between 2020 and 2021. From 2020 to 2021, the positive rates of Flu, CMV, HI and SP increased, while the positive rates of FluA and BP decreased. After the COVID-19 prevention and control measures are gradually relaxed, the positive rate of respiratory pathogens in children aged from 6 months to 6 years will increase.

Preparation and stability of pegylated poly(S-alkyl-L-homocysteine) coacervate core micelles in aqueous media.

We report development and preparation of synthetic polypeptide based, coacervate core polyelectrolyte complex micelles, PCMs, in aqueous media, which were characterized and evaluated for the encapsulation and in vitro release of a model single-stranded RNA, polyadenylic acid, poly(A). Cationic, α-helical polypeptides pegylated at their N-termini, PEG113-b-5bn and PEG113-b-5cn, were designed to form coacervate core PCMs upon mixing with multivalent anions in aqueous media. Sodium tripolyphosphate (TPP) and poly(A) were used as model multivalent anions that allowed optimization of polypeptide composition and chain length for formation of stable, nanoscale PCMs. PEG113-b-5c27 was selected for preparation of PCMs that were characterized under different environmental conditions using dynamic light scattering, atomic force microscopy and cryoelectron microscopy. The PCMs were found to efficiently encapsulate poly(A), were stable at physiologically relevant pH and solution ionic strength, and were able to release poly(A) in the presence of excess polyvalent anions. These PCMs were found to be a promising model system for further development of polypeptide based therapeutic delivery vehicles.

Quantification of changes in myocardial T1 * values with exercise cardiac MRI using a free-breathing non-electrocardiograph radial imaging.

PURPOSE: To develop and evaluate a free breathing non-electrocardiograph (ECG) myocardial T1 * mapping sequence using radial imaging to quantify the changes in myocardial T1 * between rest and exercise (T1 *reactivity ) in exercise cardiac MRI (Ex-CMR). METHODS: A free-running T1 * sequence was developed using a saturation pulse followed by three Look-Locker inversion-recovery experiments. Each Look-Locker continuously acquired data as radial trajectory using a low flip-angle spoiled gradient-echo readout. Self-navigation was performed with a temporal resolution of ∼100 ms for retrospectively extracting respiratory motion. The mid-diastole phase for every cardiac cycle was retrospectively detected on the recorded electrocardiogram signal using an empirical model. Multiple measurements were performed to obtain mean value to reduce effects from the free-breathing acquisition. Finally, data acquired at both mid-diastole and end-expiration are picked and reconstructed by a low-rank plus sparsity constraint algorithm. The performance of this sequence was evaluated by simulations, phantoms, and in vivo studies at rest and after physiological exercise. RESULTS: Numerical simulation demonstrated that changes in T1 * are related to the changes in T1 ; however, other factors such as breathing motion could influence T1 * measurements. Phantom T1 * values measured using free-running T1 * mapping sequence had good correlation with spin-echo T1 values and was insensitive to heart rate. In the Ex-CMR study, the measured T1 * reactivity was 10% immediately after exercise and declined over time. CONCLUSION: The free-running T1 * mapping sequence allows free-breathing non-ECG quantification of changes in myocardial T1 * with physiological exercise. Although, absolute myocardial T1 * value is sensitive to various confounders such as B1 and B0 inhomogeneity, quantification of its change may be useful in revealing myocardial tissue properties with exercise.

Impact of deep learning architectures on accelerated cardiac T1 mapping using MyoMapNet.

The objective of the current study was to investigate the performance of various deep learning (DL) architectures for MyoMapNet, a DL model for T1 estimation using accelerated cardiac T1 mapping from four T1 -weighted images collected after a single inversion pulse (Look-Locker 4 [LL4]). We implemented and tested three DL architectures for MyoMapNet: (a) a fully connected neural network (FC), (b) convolutional neural networks (VGG19, ResNet50), and (c) encoder-decoder networks with skip connections (ResUNet, U-Net). Modified Look-Locker inversion recovery (MOLLI) images from 749 patients at 3 T were used for training, validation, and testing. The first four T1 -weighted images from MOLLI5(3)3 and/or MOLLI4(1)3(1)2 protocols were extracted to create accelerated cardiac T1 mapping data. We also prospectively collected data from 28 subjects using MOLLI and LL4 to further evaluate model performance. Despite rigorous training, conventional VGG19 and ResNet50 models failed to produce anatomically correct T1 maps, and T1 values had significant errors. While ResUNet yielded good quality maps, it significantly underestimated T1 . Both FC and U-Net, however, yielded excellent image quality with good T1 accuracy for both native (FC/U-Net/MOLLI = 1217 ± 64/1208 ± 61/1199 ± 61 ms, all p < 0.05) and postcontrast myocardial T1 (FC/U-Net/MOLLI = 578 ± 57/567 ± 54/574 ± 55 ms, all p < 0.05). In terms of precision, the U-Net model yielded better T1 precision compared with the FC architecture (standard deviation of 61 vs. 67 ms for the myocardium for native [p < 0.05], and 31 vs. 38 ms [p < 0.05], for postcontrast). Similar findings were observed in prospectively collected LL4 data. It was concluded that U-Net and FC DL models in MyoMapNet enable fast myocardial T1 mapping using only four T1 -weighted images collected from a single LL sequence with comparable accuracy. U-Net also provides a slight improvement in precision.

Bacteroides species differentially modulate depression-like behavior via gut-brain metabolic signaling.

Gut microbiome disturbances have been widely implicated in major depressive disorder (MDD), although the identity of causal microbial species and the underlying mechanisms are yet to be fully elucidated. Here we show that Bacteroides species enriched in the gut microbiome from MDD patients differentially impact the susceptibility to depressive behaviors. Transplantation of fecal microbiome from MDD patients into antibiotic-treated mice induced anxiety and despair-like behavior and impaired hippocampal neurogenesis. Colonization of Bacteroides fragilis, Bacteroides uniformis, and, to a lesser extent, Bacteroides caccae, but not Bacteroides ovatus, recapitulated the negative effects of MDD microbiome on behavior and neurogenesis. The varying impacts of Bacteroides species were partially explained by differential alternations of tryptophan pathway metabolites and neurotransmitters along the gut-brain axis. Notably, an intensified depletion of cerebral serotonin concurred with the enhanced susceptibility to depression. Together, these findings identify select Bacteroidetes species that contribute to depression susceptibility in mice by metabolic regulation along the gut-brain axis.

Accelerated cardiac T1 mapping in four heartbeats with inline MyoMapNet: a deep learning-based T1 estimation approach.

PURPOSE: To develop and evaluate MyoMapNet, a rapid myocardial T1 mapping approach that uses fully connected neural networks (FCNN) to estimate T1 values from four T1-weighted images collected after a single inversion pulse in four heartbeats (Look-Locker, LL4). METHOD: We implemented an FCNN for MyoMapNet to estimate T1 values from a reduced number of T1-weighted images and corresponding inversion-recovery times. We studied MyoMapNet performance when trained using native, post-contrast T1, or a combination of both. We also explored the effects of number of T1-weighted images (four and five) for native T1. After rigorous training using in-vivo modified Look-Locker inversion recovery (MOLLI) T1 mapping data of 607 patients, MyoMapNet performance was evaluated using MOLLI T1 data from 61 patients by discarding the additional T1-weighted images. Subsequently, we implemented a prototype MyoMapNet and LL4 on a 3 T scanner. LL4 was used to collect T1 mapping data in 27 subjects with inline T1 map reconstruction by MyoMapNet. The resulting T1 values were compared to MOLLI. RESULTS: MyoMapNet trained using a combination of native and post-contrast T1-weighted images had excellent native and post-contrast T1 accuracy compared to MOLLI. The FCNN model using four T1-weighted images yields similar performance compared to five T1-weighted images, suggesting that four T1 weighted images may be sufficient. The inline implementation of LL4 and MyoMapNet enables successful acquisition and reconstruction of T1 maps on the scanner. Native and post-contrast myocardium T1 by MOLLI and MyoMapNet was 1170 ± 55 ms vs. 1183 ± 57 ms (P = 0.03), and 645 ± 26 ms vs. 630 ± 30 ms (P = 0.60), and native and post-contrast blood T1 was 1820 ± 29 ms vs. 1854 ± 34 ms (P = 0.14), and 508 ± 9 ms vs. 514 ± 15 ms (P = 0.02), respectively. CONCLUSION: A FCNN, trained using MOLLI data, can estimate T1 values from only four T1-weighted images. MyoMapNet enables myocardial T1 mapping in four heartbeats with similar accuracy as MOLLI with inline map reconstruction.

An inline deep learning based free-breathing ECG-free cine for exercise cardiovascular magnetic resonance.

BACKGROUND: Exercise cardiovascular magnetic resonance (Ex-CMR) is a promising stress imaging test for coronary artery disease (CAD). However, Ex-CMR requires accelerated imaging techniques that result in significant aliasing artifacts. Our goal was to develop and evaluate a free-breathing and electrocardiogram (ECG)-free real-time cine with deep learning (DL)-based radial acceleration for Ex-CMR. METHODS: A 3D (2D + time) convolutional neural network was implemented to suppress artifacts from aliased radial cine images. The network was trained using synthetic real-time radial cine images simulated using breath-hold, ECG-gated segmented Cartesian k-space data acquired at 3 T from 503 patients at rest. A prototype real-time radial sequence with acceleration rate = 12 was used to collect images with inline DL reconstruction. Performance was evaluated in 8 healthy subjects in whom only rest images were collected. Subsequently, 14 subjects (6 healthy and 8 patients with suspected CAD) were prospectively recruited for an Ex-CMR to evaluate image quality. At rest (n = 22), standard breath-hold ECG-gated Cartesian segmented cine and free-breathing ECG-free real-time radial cine images were acquired. During post-exercise stress (n = 14), only real-time radial cine images were acquired. Three readers evaluated residual artifact level in all collected images on a 4-point Likert scale (1-non-diagnostic, 2-severe, 3-moderate, 4-minimal). RESULTS: The DL model substantially suppressed artifacts in real-time radial cine images acquired at rest and during post-exercise stress. In real-time images at rest, 89.4% of scores were moderate to minimal. The mean score was 3.3 ± 0.7, representing increased (P < 0.001) artifacts compared to standard cine (3.9 ± 0.3). In real-time images during post-exercise stress, 84.6% of scores were moderate to minimal, and the mean artifact level score was 3.1 ± 0.6. Comparison of left-ventricular (LV) measures derived from standard and real-time cine at rest showed differences in LV end-diastolic volume (3.0 mL [- 11.7, 17.8], P = 0.320) that were not significantly different from zero. Differences in measures of LV end-systolic volume (7.0 mL [- 1.3, 15.3], P < 0.001) and LV ejection fraction (- 5.0% [- 11.1, 1.0], P < 0.001) were significant. Total inline reconstruction time of real-time radial images was 16.6 ms per frame. CONCLUSIONS: Our proof-of-concept study demonstrated the feasibility of inline real-time cine with DL-based radial acceleration for Ex-CMR.

Reproducibility of global and segmental myocardial strain using cine DENSE at 3 T: a multicenter cardiovascular magnetic resonance study in healthy subjects and patients with heart disease.

BACKGROUND: While multiple cardiovascular magnetic resonance (CMR) methods provide excellent reproducibility of global circumferential and global longitudinal strain, achieving highly reproducible segmental strain is more challenging. Previous single-center studies have demonstrated excellent reproducibility of displacement encoding with stimulated echoes (DENSE) segmental circumferential strain. The present study evaluated the reproducibility of DENSE for measurement of whole-slice or global circumferential (Ecc), longitudinal (Ell) and radial (Err) strain, torsion, and segmental Ecc at multiple centers. METHODS: Six centers participated and a total of 81 subjects were studied, including 60 healthy subjects and 21 patients with various types of heart disease. CMR utilized 3 T scanners, and cine DENSE images were acquired in three short-axis planes and in the four-chamber long-axis view. During one imaging session, each subject underwent two separate DENSE scans to assess inter-scan reproducibility. Each subject was taken out of the scanner and repositioned between the scans. Intra-user, inter-user-same-site, inter-user-different-site, and inter-user-Human-Deep-Learning (DL) comparisons assessed the reproducibility of different users analyzing the same data. Inter-scan comparisons assessed the reproducibility of DENSE from scan to scan. The reproducibility of whole-slice or global Ecc, Ell and Err, torsion, and segmental Ecc were quantified using Bland-Altman analysis, the coefficient of variation (CV), and the intraclass correlation coefficient (ICC). CV was considered excellent for CV ≤ 10%, good for 10% < CV ≤ 20%, fair for 20% < CV ≤ 40%, and poor for CV > 40. ICC values were considered excellent for ICC > 0.74, good for ICC 0.6 < ICC ≤ 0.74, fair for ICC 0.4 < ICC ≤ 0.59, poor for ICC < 0.4. RESULTS: Based on CV and ICC, segmental Ecc provided excellent intra-user, inter-user-same-site, inter-user-different-site, inter-user-Human-DL reproducibility and good-excellent inter-scan reproducibility. Whole-slice Ecc and global Ell provided excellent intra-user, inter-user-same-site, inter-user-different-site, inter-user-Human-DL and inter-scan reproducibility. The reproducibility of torsion was good-excellent for all comparisons. For whole-slice Err, CV was in the fair-good range, and ICC was in the good-excellent range. CONCLUSIONS: Multicenter data show that 3 T CMR DENSE provides highly reproducible whole-slice and segmental Ecc, global Ell, and torsion measurements in healthy subjects and heart disease patients.

Propofol suppresses microglial phagocytosis through the downregulation of MFG-E8.

BACKGROUND: Microglia are highly motile phagocytic cells in the healthy brain with surveillance and clearance functions. Although microglia have been shown to engulf cellular debris following brain insult, less is known about their phagocytic function in the absence of injury. Propofol can inhibit microglial activity, including phagocytosis. Milk fat globule epidermal growth factor 8 (MFG-E8), as a regulator of microglia, plays an essential role in the phagocytic process. However, whether MFG-E8 affects the alteration of phagocytosis by propofol remains unknown. METHODS: Microglial BV2 cells were treated with propofol, with or without MFG-E8. Phagocytosis of latex beads was evaluated by flow cytometry and immunofluorescence. MFG-E8, p-AMPK, AMPK, p-Src, and Src levels were assessed by western blot analysis. Compound C (AMPK inhibitor) and dasatinib (Src inhibitor) were applied to determine the roles of AMPK and Src in microglial phagocytosis under propofol treatment. RESULTS: The phagocytic ability of microglia was significantly decreased after propofol treatment for 4 h (P < 0.05). MFG-E8 production was inhibited by propofol in a concentration- and time-dependent manner (P < 0.05). Preadministration of MFG-E8 dose-dependently (from 10 to 100 ng/ml) reversed the suppression of phagocytosis by propofol (P < 0.05). Furthermore, the decline in p-AMPK and p-Src levels induced by propofol intervention was reversed by MFG-E8 activation (P < 0.05). Administration of compound C (AMPK inhibitor) and dasatinib (Src inhibitor) to microglia blocked the trend of enhanced phagocytosis induced by MFG-E8 (P < 0.05). CONCLUSIONS: These findings reveal the intermediate role of MFG-E8 between propofol and microglial phagocytic activity. Moreover, MFG-E8 may reverse the suppression of phagocytosis induced by propofol through the regulation of the AMPK and Src signaling pathways.

Free-breathing whole-heart multi-slice myocardial T1 mapping in 2 minutes.

PURPOSE: To develop and validate a saturation-delay-inversion recovery preparation, slice tracking and multi-slice based sequence for measuring whole-heart native T1 . METHOD: The proposed free-breathing sequence performs T1 mapping of multiple left-ventricular slices by slice-interleaved acquisition to collect 10 electrocardiogram-triggered single-shot slice-selective images for each slice. A saturation-delay-inversion recovery pulse is used for T1 preparation. Prospective slice tracking by the diaphragm navigator and retrospective registration are used to reduce through-plane and in-plane motion, respectively. The proposed sequence was validated in both phantom and human subjects (12 healthy subjects and 15 patients who were referred for a clinical cardiac MR exam) and compared with saturation recovery single-shot acquisition (SASHA) and modified Look-Locker inversion recovery (MOLLI). RESULTS: Phantom T1 measured by the proposed sequence had excellent agreement (R2  = 0.99) with the ground-truth T1 and was insensitive to heart rate. In both healthy subjects and patients, the proposed sequence yielded nine left-ventricular T1 maps per volume in less than 2 minutes (healthy volunteers: 1.8 ± 0.4 minutes; patients: 1.9 ± 0.2 minutes). The average T1 of whole left ventricle for all healthy subjects and patients were 1560 ± 61 and 1535 ± 49 ms by SASHA, 1208 ± 42 and 1233 ± 56 ms by MOLLI5(3)3, and 1397 ± 34 and 1433 ± 56 ms by the proposed sequence, respectively. The corresponding coefficient of variation of T1 were 6.2 ± 1.4% and 5.8 ± 1.6%, 5.3 ± 1.1% and 5.1 ± 0.8%, and 4.9 ± 0.8% and 4.5 ± 0.8%, respectively. CONCLUSION: The proposed sequence enables quantification of whole heart T1 with good accuracy and precision in less than 2 minutes during free breathing.

Artifact reduction in free-breathing, free-running myocardial perfusion imaging with interleaved non-selective RF excitations.

PURPOSE: To reduce inflow and motion artifacts in free-breathing, free-running, steady-state spoiled gradient echo T1 -weighted (SPGR) myocardial perfusion imaging. METHOD: Unsaturated spins from inflowing blood or out-of-plane motion cause flashing artifacts in free-running SPGR myocardial perfusion. During free-running SPGR, 1 non-selective RF excitation was added after every 3 slice-selective RF excitations to suppress inflow artifacts by forcing magnetization in neighboring regions to steady-state. Bloch simulations and phantom experiments were performed to evaluate the impact of the flip angle and non-selective RF frequency on inflowing spins and tissue contrast. Free-running perfusion with (n = 11) interleaved non-selective RF or without (n = 11) were studied in 22 subjects (age = 60.2 ± 14.3 years, 11 male). Perfusion images were graded on a 5-point Likert scale for conspicuity of wall enhancement, inflow/motion artifact, and streaking artifact and compared using Wilcoxon sum-rank testing. RESULT: Numeric simulation showed that 1 non-selective RF excitation applied after every 3 slice-selective RF excitations produced superior out-of-plane signal suppression compared to 1 non-selective RF excitation applied after every 6 or 9 slice-selective RF excitations. In vitro experiments showed that a 30° flip angle produced near-optimal myocardial contrast. In vivo experiments demonstrated that the addition of interleaved non-selective RF significantly (P < .01) improved conspicuity of wall enhancement (mean score = 4.4 vs. 3.2) and reduced inflow/motion (mean score = 4.5 vs. 2.5) and streaking (mean score = 3.9 vs. 2.4) artifacts. CONCLUSION: Non-selective RF excitations interleaved between slice-selective excitations can reduce image artifacts in free-breathing, ungated perfusion images. Further studies are warranted to assess the diagnostic accuracy of the proposed solution for evaluating myocardial ischemia.

Free-breathing simultaneous myocardial T1 and T2 mapping with whole left ventricle coverage.

PURPOSE: To develop a free-breathing sequence, that is, Multislice Joint T1 -T2 , for simultaneous measurement of myocardial T1 and T2 for multiple slices to achieve whole left-ventricular coverage. METHODS: Multislice Joint T1 -T2 adopts slice-interleaved acquisition to collect 10 single-shot electrocardiogram-triggered images for each slice prepared by saturation and T2 preparation to simultaneously estimate myocardial T1 and T2 and achieve whole left-ventricular coverage. Prospective slice-tracking using a respiratory navigator and retrospective image registration are used to reduce through-plane and in-plane motion, respectively. Multislice Joint T1 -T2 was validated through numerical simulations and phantom and in vivo experiments, and compared with saturation-recovery single-shot acquisition and T2 -prepared balanced Steady-State Free Precession (T2 -prep SSFP) sequences. RESULTS: Phantom T1 and T2 from Multislice Joint T1 -T2 had good accuracy and precision, and were insensitive to heart rate. Multislice Joint T1 -T2 yielded T1 and T2 maps of nine left-ventricular slices in 1.4 minutes. The mean left-ventricular T1 difference between saturation-recovery single-shot acquisition and Multislice Joint T1 -T2 across healthy subjects and patients was 191 ms (1564 ± 60 ms versus 1373 ± 50 ms; P < .05) and 111 ms (1535 ± 49 ms vs 1423 ± 49 ms; P < .05), respectively. The mean difference in left-ventricular T2 between T2 -prep SSFP and Multislice Joint T1 -T2 across healthy subjects and patients was -6.3 ms (42.4 ± 1.4 ms vs 48.7 ± 2.5; P < .05) and -5.7 ms (41.6 ± 2.5 ms vs 47.3 ± 2.7; P < .05), respectively. CONCLUSION: Multislice Joint T1 -T2 enables quantification of whole left-ventricular T1 and T2 during free breathing within a clinically feasible scan time of less than 2 minutes.

Multi-domain convolutional neural network (MD-CNN) for radial reconstruction of dynamic cardiac MRI.

PURPOSE: Cardiac MR cine imaging allows accurate and reproducible assessment of cardiac function. However, its long scan time not only limits the spatial and temporal resolutions but is challenging in patients with breath-holding difficulty or non-sinus rhythms. To reduce scan time, we propose a multi-domain convolutional neural network (MD-CNN) for fast reconstruction of highly undersampled radial cine images. METHODS: MD-CNN is a complex-valued network that processes MR data in k-space and image domains via k-space interpolation and image-domain subnetworks for residual artifact suppression. MD-CNN exploits spatio-temporal correlations across timeframes and multi-coil redundancies to enable high acceleration. Radial cine data were prospectively collected in 108 subjects (50 ± 17 y, 72 males) using retrospective-gated acquisition with 80%:20% split for training/testing. Images were reconstructed by MD-CNN and k-t Radial Sparse-Sense(kt-RASPS) using an undersampled dataset (14 of 196 acquired views; relative acceleration rate = 14). MD-CNN images were evaluated quantitatively using mean-squared-error (MSE) and structural similarity index (SSIM) relative to reference images, and qualitatively by three independent readers for left ventricular (LV) border sharpness and temporal fidelity using 5-point Likert-scale (1-non-diagnostic, 2-poor, 3-fair, 4-good, and 5-excellent). RESULTS: MD-CNN showed improved MSE and SSIM compared to kt-RASPS (0.11 ± 0.10 vs. 0.61 ± 0.51, and 0.87 ± 0.07 vs. 0.72 ± 0.07, respectively; P < .01). Qualitatively, MD-CCN significantly outperformed kt-RASPS in LV border sharpness (3.87 ± 0.66 vs. 2.71 ± 0.58 at end-diastole, and 3.57 ± 0.6 vs. 2.56 ± 0.6 at end-systole, respectively; P < .01) and temporal fidelity (3.27 ± 0.65 vs. 2.59 ± 0.59; P < .01). CONCLUSION: MD-CNN reduces the scan time of cine imaging by a factor of 23.3 and provides superior image quality compared to kt-RASPS.

Highly accelerated free-breathing real-time phase contrast cardiovascular MRI via complex-difference deep learning.

PURPOSE: To develop and evaluate a real-time phase contrast (PC) MRI protocol via complex-difference deep learning (DL) framework. METHODS: DL used two 3D U-nets to separately filter aliasing artifact from radial real-time velocity-compensated and complex-difference images. U-nets were trained with synthetic real-time PC generated from electrocardiograph (ECG) -gated, breath-hold, segmented PC (ECG-gated segmented PC) acquired at the ascending aorta of 510 patients. In 21 patients, free-breathing, ungated real-time (acceleration rate = 28.8) and ECG-gated segmented (acceleration rate = 2) PC were prospectively acquired at the ascending aorta. Hemodynamic parameters (cardiac output [CO], stroke volume [SV], and mean velocity at peak systole [peak mean velocity]) were measured for ECG-gated segmented and DL-filtered synthetic real-time PC and compared using Bland-Altman and linear regression analyses. Additionally, hemodynamic parameters were quantified from DL-filtered, compressed-sensing (CS) -reconstructed, and gridding reconstructed prospective real-time PC and compared to ECG-gated segmented PC. RESULTS: Synthetic real-time PC with DL showed strong correlation (R > 0.98) and good agreement with ECG-gated segmented PC for quantified hemodynamic parameters (mean-difference: CO = -0.3 L/min, SV = -4.3 mL, peak mean velocity = -2.3 cm/s). On average, DL required 0.39 s/frame to filter prospective real-time PC, which was 4.6-fold faster than CS. Compared to CS, DL showed superior correlation, tighter limits of agreement (LOAs), better bias for peak mean velocity, and worse bias for CO and SV. Compared to gridding, DL showed similar correlation, tighter LOAs for CO and SV, similar bias for CO, and worse bias for SV and peak mean velocity. CONCLUSION: The complex-difference DL framework accelerated real-time PC-MRI by nearly 28-fold, enabling rapid free-running real-time assessment of flow hemodynamics.

Exploring the stability of inhibitor binding to SIK2 using molecular dynamics simulation and binding free energy calculation.

Salt inducible kinase 2 (SIK2) is a calcium/calmodulin-dependent protein kinase-like kinase that is implicated in a variety of biological phenomena, including cellular metabolism, growth, and apoptosis. SIK2 is the key target for various cancers, including ovarian, breast, prostate, and lung cancers. Although potent inhibitors of SIK2 are being developed, their binding stability and functional role are not presently known. In this work, we studied the detailed interactions between SIK2 and four of its inhibitors, HG-9-91-01, KIN112, MRT67307, and MRT199665, using molecular docking, molecular dynamics simulation, binding free energy calculation, and interaction fingerprint analysis. Intermolecular interactions revealed that HG-9-91-01 and KIN112 have stronger interactions with SIK2 than those of MRT199665 and MRT67307. The key residues involved in binding with SIK2 are conserved among all four inhibitors. Our results explain the detailed interaction of SIK2 with its inhibitors at the molecular level, thus paving the way for the development of targeted efficient anti-cancer drugs.

Mutation in the intracellular chloride channel CLCC1 associated with autosomal recessive retinitis pigmentosa.

We identified a homozygous missense alteration (c.75C>A, p.D25E) in CLCC1, encoding a presumptive intracellular chloride channel highly expressed in the retina, associated with autosomal recessive retinitis pigmentosa (arRP) in eight consanguineous families of Pakistani descent. The p.D25E alteration decreased CLCC1 channel function accompanied by accumulation of mutant protein in granules within the ER lumen, while siRNA knockdown of CLCC1 mRNA induced apoptosis in cultured ARPE-19 cells. TALEN KO in zebrafish was lethal 11 days post fertilization. The depressed electroretinogram (ERG) cone response and cone spectral sensitivity of 5 dpf KO zebrafish and reduced eye size, retinal thickness, and expression of rod and cone opsins could be rescued by injection of wild type CLCC1 mRNA. Clcc1+/- KO mice showed decreased ERGs and photoreceptor number. Together these results strongly suggest that intracellular chloride transport by CLCC1 is a critical process in maintaining retinal integrity, and CLCC1 is crucial for survival and function of retinal cells.

Non-Cartesian slice-GRAPPA and slice-SPIRiT reconstruction methods for multiband spiral cardiac MRI.

PURPOSE: Spiral MRI has advantages for cardiac imaging, and multiband (MB) spiral MRI of the heart shows promise. However, current reconstruction methods for MB spiral imaging have limitations. We sought to develop improved reconstruction methods for MB spiral cardiac MRI. METHODS: Two reconstruction methods were developed. The first is non-Cartesian slice-GRAPPA (NCSG), which uses phase demodulation and gridding operations before application of a Cartesian slice-separating kernel. The second method, slice-SPIRiT, formulates the reconstruction as a minimization problem that enforces in-plane coil consistency and consistency with the acquired MB data, and uses through-plane coil sensitivity information in the iterative solution. These methods were compared with conjugate-gradient SENSE in phantoms and volunteers. Temporal alternation of CAIPIRINHA (controlled aliasing in parallel imaging results in higher acceleration) phase and the use of a temporal filter were also investigated. RESULTS: Phantom experiments with 3 simultaneous slices (MB = 3) showed that mean artifact power was highest for conjugate-gradient SENSE, lower for NCSG, and lowest for slice-SPIRiT. For volunteer cine imaging (MB = 3, N = 5), the artifact power was 0.182 ± 0.037, 0.148 ± 0.036, and 0.139 ± 0.034 for conjugate-gradient SENSE, NCSG, and slice-SPIRiT, respectively (P < .05, analysis of variance). Temporal alternation of CAIPIRINHA reduced artifacts for both NCSG and slice-SPIRiT. CONCLUSION: The NCSG and slice-SPIRiT methods provide more accurate reconstructions for MB spiral cine imaging compared with conjugate-gradient SENSE. These methods hold promise for non-Cartesian MB imaging.

Flavonoids with Inhibitory Effects on NLRP3 Inflammasome Activation from Millettia velutina.

Eight new flavonoids, including two ß-hydroxy/methoxychalcones, velutones A and B (1 and 2), two 1,3-diarylpropan-1-ols, velutols C and D (3 and 4), a dihydroxychalcone, velutone E (5), a chalcone, velutone F (6), a furanoflavanone, velutone G (7), and a furanoflavonol, velutone H (8), and 14 known compounds were isolated from Millettia velutina. Their structures were determined by high-resolution electrospray ionisation mass spectrometry (HR-ESIMS) and spectroscopic data analyses and time-dependent density functional theory electronic circular dichroism (TD-DFT-ECD) calculations. Among the isolated constituents, compound 6 exhibited the most potent inhibitory effect (IC50: 1.3 µM) against nigericin-induced IL-1ß release in THP-1 cells. The initial mechanism of action study revealed that compound 6 suppressed NLRP3 inflammasome activation via blocking ASC oligomerization without affecting the priming step, which subsequently inhibited caspase-1 activation and IL-1ß secretion. Most importantly, compound 6 exerted potent protective effects in the LPS-induced septic shock mice model by improving the survival rate of mice and suppressing serum IL-1ß release. These results demonstrated that compound 6 had the potential to be developed as a broad-spectrum NLRP3 inflammasome inhibitor for the treatment of NLRP3-related disease.