FISH-Flow to quantify nascent and mature ribosomal RNA in mouse and human cells.

FISH-Flow (fluorescence in situ hybridization-flow cytometry) involves hybridizing fluorescent oligos to RNA and quantifying fluorescence at a single-cell level using flow cytometry. Here, we present a FISH-Flow protocol to quantify nascent 47S and mature 18S and 28S rRNAs in mouse and human cells, including rRNA quantification across cell cycle stages using DNA staining. We describe steps for cell preparation, hybridization of fluorescent probes against rRNA, and DNA staining. We then detail procedures for flow cytometry and data analysis. For complete details on the use and execution of this protocol, please refer to Antony et al. (2022).1.

Evaluation of a compact 3 T MRI scanner for patients with implanted devices.

Access to high-quality MR exams is severely limited for patients with some implanted devices due to labeled MR safety conditions, but small-bore systems can overcome this limitation. For example, a compact 3 T MR scanner (C3T) with high-performance gradients can acquire exams of the head, extremities, and infants. Because of its reduced bore size and the patient being advanced only partially into the bore, the associated electromagnetic (EM) fields drop off rapidly caudal to the head, compared to whole-body systems. Therefore, some patients with MR conditional implanted devices can safely receive 3 T brain exams on the C3T using its strong gradients and a multiple-channel receive coil, while a corresponding exam on whole-body MR is precluded. The purpose of this study is to evaluate the performance of a small-bore scanner for subjects with MR conditional spinal or sacral nerve stimulators, or abandoned cardiac implantable electronic device (CIED) leads. The spatial dependence of specific absorption rate (SAR) on the C3T was compared to whole-body scanners. A device assessment tool was developed and applied to evaluate MR safety individually on the C3T for 12 subjects with implanted devices or abandoned CIED leads. Once MR safety was established, the subjects received a C3T brain exam along with their clinical, 1.5 T exam. The resulting images were graded by three board-certified neuroradiologists. The C3T exams were well-tolerated with no adverse events, and significantly outperformed the whole-body 1.5 T exams in terms of overall image quality.

Improved Resting-State Functional MRI Using Multi-Echo Echo-Planar Imaging on a Compact 3T MRI Scanner with High-Performance Gradients.

In blood-oxygen-level-dependent (BOLD)-based resting-state functional (RS-fMRI) studies, usage of multi-echo echo-planar-imaging (ME-EPI) is limited due to unacceptable late echo times when high spatial resolution is used. Equipped with high-performance gradients, the compact 3T MRI system (C3T) enables a three-echo whole-brain ME-EPI protocol with smaller than 2.5 mm isotropic voxel and shorter than 1 s repetition time, as required in landmark fMRI studies. The performance of the ME-EPI was comprehensively evaluated with signal variance reduction and region-of-interest-, seed- and independent-component-analysis-based functional connectivity analyses and compared with a counterpart of single-echo EPI with the shortest TR possible. Through the multi-echo combination, the thermal noise level is reduced. Functional connectivity, as well as signal intensity, are recovered in the medial orbital sulcus and anterior transverse collateral sulcus in ME-EPI. It is demonstrated that ME-EPI provides superior sensitivity and accuracy for detecting functional connectivity and/or brain networks in comparison with single-echo EPI. In conclusion, the high-performance gradient enabled high-spatial-temporal resolution ME-EPI would be the method of choice for RS-fMRI study on the C3T.

Enhanced clinical task-based fMRI metrics through locally low-rank denoising of complex-valued data.

OBJECTIVE: This study investigates a locally low-rank (LLR) denoising algorithm applied to source images from a clinical task-based functional MRI (fMRI) exam before post-processing for improving statistical confidence of task-based activation maps. METHODS: Task-based motor and language fMRI was obtained in eleven healthy volunteers under an IRB approved protocol. LLR denoising was then applied to raw complex-valued image data before fMRI processing. Activation maps generated from conventional non-denoised (control) data were compared with maps derived from LLR-denoised image data. Four board-certified neuroradiologists completed consensus assessment of activation maps; region-specific and aggregate motor and language consensus thresholds were then compared with nonparametric statistical tests. Additional evaluation included retrospective truncation of exam data without and with LLR denoising; a ROI-based analysis tracked t-statistics and temporal SNR (tSNR) as scan durations decreased. A test-retest assessment was performed; retest data were matched with initial test data and compared for one subject. RESULTS: fMRI activation maps generated from LLR-denoised data predominantly exhibited statistically significant (p = 4.88×10-4 to p = 0.042; one p = 0.062) increases in consensus t-statistic thresholds for motor and language activation maps. Following data truncation, LLR data showed task-specific increases in t-statistics and tSNR respectively exceeding 20 and 50% compared to control. LLR denoising enabled truncation of exam durations while preserving cluster volumes at fixed thresholds. Test-retest showed variable activation with LLR data thresholded higher in matching initial test data. CONCLUSION: LLR denoising affords robust increases in t-statistics on fMRI activation maps compared to routine processing, and offers potential for reduced scan duration while preserving map quality.

Sequence-specific destabilization of azurin by tetramethylguanidinium-dipeptide ionic liquids.

The thermal unfolding of the copper redox protein azurin was studied in the presence of four different dipeptide-based ionic liquids (ILs) utilizing tetramethylguanidinium as the cation. The four dipeptides have different sequences including the amino acids Ser and Asp: TMG-AspAsp, TMG-SerSer, TMG-SerAsp, and TMG-AspSer. Thermal unfolding curves generated from temperature-dependent fluorescence spectroscopy experiments showed that TMG-AspAsp and TMG-SerSer have minor destabilizing effects on the protein while TMG-AspSer and TMG-SerAsp strongly destabilize azurin. Red-shifted fluorescence signatures in the 25 °C correlate with the observed protein destabilization in the solutions with TMG-AspSer and TMG-SerAsp. These signals could correspond to interactions between the Asp residue in the dipeptide and the azurin Trp residue in the unfolded state. These results, supported by appropriate control experiments, suggest that dipeptide sequence-specific interactions lead to selective protein destabilization and motivate further studies of TMG-dipeptide ILs.

Improved Brain MR Imaging from a Compact, Lightweight 3T Scanner with High-Performance Gradients.

BACKGROUND: A low-cryogen, compact 3T (C3T) MRI scanner with high-performance gradients capable of simultaneously achieving 80 mT/m gradient amplitude and 700 T/m/second slew rate has been in use to study research patients since March 2016 but has not been implemented in the clinical practice. PURPOSE: To compare head MRI examinations obtained with the C3T system and a conventional whole-body 3T (WB3T) scanner in seven parameters across five commonly used brain imaging sequences. STUDY TYPE: Prospective. SUBJECTS: Thirty patients with a clinically indicated head MRI. SEQUENCE: 3T; T1 FLAIR, T1 MP-RAGE, 3D T2 FLAIR, T2 FSE, and DWI. ASSESSMENT: All patients tolerated the scans well. Three board-certified neuroradiologists scored the comparative quality of C3T and WB3T images in blinded fashion using a five-point Likert scale in terms of: signal-to-noise ratio, lesion conspicuity, motion artifact, gray/white matter contrast, cerebellar folia, susceptibility artifact, and overall quality. STATISTICAL TEST: Left-sided, right-sided, and two-sided Wilcoxon signed rank test; Fisher's method. A P value <0.05 was considered statistically significant. RESULTS: The C3T system performed better than the WB3T in virtually all comparisons, except for motion artifacts for the T1 FLAIR and T1 MP-RAGE sequences, where the WB3T system was deemed better. When combining all sequences together, the C3T system outperformed the WB3T system in all image quality parameters evaluated, except for motion artifact (P = 0.13). DATA CONCLUSION: The C3T scanner provided better overall image quality for all sequences, and performed better in all individual categories, except for motion artifact on the T1 FLAIR and T1 MP-RAGE. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY STAGE: 1.

Application of Adaptive Image Receive Coil Technology for Whole-Brain Imaging.

OBJECTIVE. The Adaptive Image Receive (AIR) radiofrequency coil is an emergent technology that is lightweight and flexible and exhibits electrical characteristics that overcome many of the limitations of traditional rigid coil designs. The purpose of this study was to apply the AIR coil for whole-brain imaging and compare the performance of a prototype AIR coil array with the performance of conventional head coils. SUBJECTS AND METHODS. A phantom and 15 healthy adult participants were imaged. A prototype 16-channel head AIR coil was compared with conventional 8-and 32-channel head coils using clinically available MRI sequences. During consensus review, two board-certified neuroradiologists graded the AIR coil compared with an 8-channel coil and a 32-channel coil on a 5-point ordinal scale in multiple categories. One- and two-sided Wilcoxon signed rank tests were performed. Noise covariance matrices and geometry factor (g-factor) maps were calculated. RESULTS. The signal-to-noise ratio, structural sharpness, and overall image quality scores of the prototype 16-channel AIR coil were better than those of the 8-channel coil but were not as good as those of the 32-channel coil. Noise covariance matrices showed stable performance of the AIR coil across participants. The median g-factors for the 16-channel AIR coil were, overall, less than those of the 8-channel coil but were greater than those of the 32-channel coil. CONCLUSION. On average, the prototype 16-channel head AIR coil outperformed a conventional 8-channel head coil but did not perform as well as a conventional 32-channel head coil. This study shows the feasibility of the novel AIR coil technology for imaging the brain and provides insight for future coil design improvements.

Development and validation of a potent and specific inhibitor for the CLC-2 chloride channel.

CLC-2 is a voltage-gated chloride channel that is widely expressed in mammalian tissues. In the central nervous system, CLC-2 appears in neurons and glia. Studies to define how this channel contributes to normal and pathophysiological function in the central nervous system raise questions that remain unresolved, in part due to the absence of precise pharmacological tools for modulating CLC-2 activity. Herein, we describe the development and optimization of AK-42, a specific small-molecule inhibitor of CLC-2 with nanomolar potency (IC50 = 17 ± 1 nM). AK-42 displays unprecedented selectivity (>1,000-fold) over CLC-1, the closest CLC-2 homolog, and exhibits no off-target engagement against a panel of 61 common channels, receptors, and transporters expressed in brain tissue. Computational docking, validated by mutagenesis and kinetic studies, indicates that AK-42 binds to an extracellular vestibule above the channel pore. In electrophysiological recordings of mouse CA1 hippocampal pyramidal neurons, AK-42 acutely and reversibly inhibits CLC-2 currents; no effect on current is observed on brain slices taken from CLC-2 knockout mice. These results establish AK-42 as a powerful tool for investigating CLC-2 neurophysiology.

The benefit of high-performance gradients on echo planar imaging for BOLD-based resting-state functional MRI.

Improved gradient performance in an MRI system reduces distortion in echo planar imaging (EPI), which has been a key imaging method for functional studies. A lightweight, low-cryogen compact 3T MRI scanner (C3T) is capable of achieving 80 mT m-1 gradient amplitude with 700 T m-1 s-1 slew rate, in comparison with a conventional whole-body 3T MRI scanner (WB3T, 50 mT m-1 with 200 T m-1 s-1). We investigated benefits of the high-performance gradients in a high-spatial-resolution (1.5 mm isotropic) functional MRI study. Reduced echo spacing in the EPI pulse sequence inherently leads to less severe geometric distortion, which provided higher accuracy than with WB3T for registration between EPI and anatomical images. The cortical coverage of C3T datasets was improved by more accurate signal depiction (i.e. less dropout or pile-up). Resting-state functional analysis results showed that greater magnitude and extent in functional connectivity (FC) for the C3T than the WB3T when the selected seed region is susceptible to distortions, while the FC matrix for well-known brain networks showed little difference between the two scanners. This shows that the improved quality in EPI is particularly valuable for studying certain brain regions typically obscured by severe distortion.

Reducing PNS with minimal performance penalties via simple pulse sequence modifications on a high-performance compact 3T scanner.

One of the major concerns associated with high-performance gradients is peripheral nerve stimulation (PNS) of the subject during MRI exams. Since the installation, more than 680 volunteer subjects (patients and controls) have been scanned on a compact 3 T MRI system with high-performance gradients, capable of 80 mT m-1 gradient amplitude and 700 T m-1 s-1 slew rate simultaneously. Despite PNS concerns associated with the high-performance gradients, due to the smaller physical dimensions of the gradient coils, minimal or no PNS sensation was reported with most pulse sequences. The exception was PNS reported by only five of 252 subjects (about 2%) scanned with a specific 3D fast spin echo pulse sequence (3DFLAIR). Rather than derating the entire system performance across all pulse sequences and all gradient lobes, we addressed reported PNS effect with a simple and specific modification to the targeted lobes of the problematic pulse sequence. in addition, the PNS convolutional model was adapted to predict sequence-specific PNS threshold level and its reduction after derating. The effectiveness of the targeted pulse sequence modification was demonstrated by successfully re-scanning four of the subjects who previously reported PNS sensations without further reported PNS. The pulse sequence modification did not result in noticeable degradation of image quality or substantial increase in scan time. The results demonstrated that PNS was rarely reported on the compact 3 T, and when it was, utilizing a specific modification of the gradient waveform causing PNS was an effective strategy, rather than derating the performance of the entire gradient system.

Distortion-free imaging: A double encoding method (DIADEM) combined with multiband imaging for rapid distortion-free high-resolution diffusion imaging on a compact 3T with high-performance gradients.

BACKGROUND: Distortion-free, high-resolution diffusion imaging using DIADEM (Distortion-free Imaging: A Double Encoding Method), proposed recently, has great potential for clinical applications. However, it can suffer from prolonged scan times and its reliability for quantitative diffusion imaging has not been evaluated. PURPOSE: To investigate the clinical feasibility of DIADEM-based high-resolution diffusion imaging on a novel compact 3T (C3T) by evaluating the reliability of quantitative diffusion measurements and utilizing both the high-performance gradients (80 mT/m, 700 T/m/s) and the sequence optimization with the navigator acquisition window reduction and simultaneous multislice (multiband) imaging. STUDY TYPE: Prospective feasibility study. PHANTOM/SUBJECTS: Diffusion quality control phantom scans to evaluate the reliability of quantitative diffusion measurements; 36 normal control scans for B0 -field mapping; six healthy and two patient subject scans with a brain tumor for comparisons of diffusion and anatomical imaging. FIELD STRENGTH/SEQUENCE: 3T; the standard single-shot echo-planar-imaging (EPI), multishot DIADEM diffusion, and anatomical (2D-FSE [fast-spin-echo], 2D-FLAIR [fluid-attenuated-inversion-recovery], and 3D-MPRAGE [magnetization prepared rapid acquisition gradient echo]) imaging. ASSESSMENT: The scan time reduction, the reliability of quantitative diffusion measurements, and the clinical efficacy for high-resolution diffusion imaging in healthy control and brain tumor volunteers. STATISTICAL TEST: Bland-Altman analysis. RESULTS: The scan time for high in-plane (0.86 mm2 ) resolution, distortion-free, and whole brain diffusion imaging were reduced from 10 to 5 minutes with the sequence optimizations. All of the mean apparent diffusion coefficient (ADC) values in phantom were within the 95% confidence interval in the Bland-Altman plot. The proposed acquisition with a total off-resonance coverage of 597.2 Hz wider than the expected bandwidth of 500 Hz in human brain could yield a distortion-free image without foldover artifacts. Compared with EPI, therefore, this approach allowed direct image matching with the anatomical images and enabled improved delineation of the tumor boundaries. DATA CONCLUSION: The proposed high-resolution diffusion imaging approach is clinically feasible on C3T due to a combination of hardware and sequence improvements. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 1 J. Magn. Reson. Imaging 2020;51:296-310.

The effect of spiral trajectory correction on pseudo-continuous arterial spin labeling with high-performance gradients on a compact 3T scanner.

PURPOSE: To demonstrate the feasibility of pseudo-continuous arterial-spin-labeled (pCASL) imaging with 3D fast-spin-echo stack-of-spirals on a compact 3T scanner (C3T), to perform trajectory correction for eddy-current-induced deviations in the spiral readout of pCASL imaging, and to assess the correction effect on perfusion-related images with high-performance gradients (80 mT/m, 700T/m/s) of the C3T. METHODS: To track eddy-current-induced artifacts with Archimedean spiral readout, the spiral readout in pCASL imaging was performed with 5 different peak gradient slew rate (Smax ) values ranging from 70 to 500 T/m/s. The trajectory for each Smax was measured using a dynamic field camera and applied in a density-compensated gridding image reconstruction in addition to the nominal trajectory. The effect of the trajectory correction was assessed with perfusion-weighted (ΔM) images and proton-density-weighted images as well as cerebral blood flow (CBF) maps, obtained from 10 healthy volunteers. RESULTS: Blurring artifact on ΔM images was mitigated by the trajectory correction. CBF values on the left and right calcarine cortices showed no significant difference after correction. Also, the signal-to-noise ratio of ΔM images improved, on average, by 7.6% after correction (P < .001). The greatest improvement of 12.1% on ΔM images was achieved with a spiral readout using Smax of 300~400 T/m/s. CONCLUSION: Eddy currents can cause spiral trajectory deviation, which leads to deformation of the CBF map even in cases of low value Smax . The trajectory correction for spiral-readout-based pCASL produces more reliable results for perfusion imaging. These results suggest that pCASL is feasible on C3T with high-performance gradients.

Population extinctions driven by climate change, population size, and time since observation may make rare species databases inaccurate.

Loss of biological diversity through population extinctions is a global phenomenon that threatens many ecosystems. Managers often rely on databases of rare species locations to plan land use actions and conserve at-risk taxa, so it is crucial that the information they contain is accurate and dependable. However, small population sizes, long gaps between surveys, and climate change may be leading to undetected extinctions of many populations. We used repeated survey records for a rare but widespread orchid, Cypripedium fasciculatum (clustered lady's slipper), to model population extinction risk based on elevation, population size, and time between observations. Population size and elevation were negatively associated with extinction, while extinction probability increased with time between observations. We interpret population losses at low elevations as a potential signal of climate change impacts. We used this model to estimate the probability of persistence of populations across California and Oregon, and found that 39%-52% of the 2415 populations reported in databases from this region are likely extinct. Managers should be aware that the number of populations of rare species in their databases is potentially an overestimate, and consider resurveying these populations to document their presence and condition, with priority given to older reports of small populations, especially those at low elevations or in other areas with high vulnerability to climate or land cover change.

Disruption of GpI mGluR-Dependent Cav2.3 Translation in a Mouse Model of Fragile X Syndrome.

Fragile X syndrome (FXS) is an inherited intellectual impairment that results from the loss of fragile X mental retardation protein (FMRP), an mRNA binding protein that regulates mRNA translation at synapses. The absence of FMRP leads to neuronal and circuit-level hyperexcitability that is thought to arise from the aberrant expression and activity of voltage-gated ion channels, although the identification and characterization of these ion channels have been limited. Here, we show that FMRP binds the mRNA of the R-type voltage-gated calcium channel Cav2.3 in mouse brain synaptoneurosomes and represses Cav2.3 translation under basal conditions. Consequently, in hippocampal neurons from male and female FMRP KO mice, we find enhanced Cav2.3 protein expression by western blotting and abnormally large R currents in whole-cell voltage-clamp recordings. In agreement with previous studies showing that FMRP couples Group I metabotropic glutamate receptor (GpI mGluR) signaling to protein translation, we find that GpI mGluR stimulation results in increased Cav2.3 translation and R current in hippocampal neurons which is disrupted in FMRP KO mice. Thus, FMRP serves as a key translational regulator of Cav2.3 expression under basal conditions and in response to GpI mGluR stimulation. Loss of regulated Cav2.3 expression could underlie the neuronal hyperactivity and aberrant calcium spiking in FMRP KO mice and contribute to FXS, potentially serving as a novel target for future therapeutic strategies.SIGNIFICANCE STATEMENT Patients with fragile X syndrome (FXS) exhibit signs of neuronal and circuit hyperexcitability, including anxiety and hyperactive behavior, attention deficit disorder, and seizures. FXS is caused by the loss of fragile X mental retardation protein (FMRP), an mRNA binding protein, and the neuronal hyperexcitability observed in the absence of FMRP likely results from its ability to regulate the expression and activity of voltage-gated ion channels. Here we find that FMRP serves as a key translational regulator of the voltage-gated calcium channel Cav2.3 under basal conditions and following activity. Cav2.3 impacts cellular excitability and calcium signaling, and the alterations in channel translation and expression observed in the absence of FMRP could contribute to the neuronal hyperactivity that underlies FXS.

On the outside looking in: Finding a place for managed alcohol programs in the harm reduction movement.

Alcohol policy in North America is dominated by moderation and abstinence-based modalities that focus on controlling population-level alcohol consumption and modifying individual consumption patterns to prevent and reduce alcohol-related harms. However, conventional alcohol policies and interventions do not adequately address harms associated with high-risk drinking among individuals experiencing severe alcohol use disorder (AUD) and structural vulnerability such as poverty and homelessness. In this commentary we address this gap in alcohol harm reduction, and highlight the lack of, and distinct need for, alcohol-specific harm reduction for people experiencing structural vulnerability and severe AUD. These individuals, doubly impacted by structural oppression and severe AUD, engage in various high-risk drinking practices that contribute to a unique set of harms that conventional abstinence-based treatments and interventions fail to adequately attend to. Managed alcohol programs (MAPs) have been established to address these multiple intersecting harms, and though gaining momentum across Canada, have had a hard time finding their place within the harm reduction movement. We illustrate how MAPs play a crucial role in the harm reduction movement in their ability to not only address high-risk drinking practices among structurally marginalized individuals, but to respond to harms associated with broader structural inequities such as poverty and homelessness.

Lightweight, compact, and high-performance 3T MR system for imaging the brain and extremities.

PURPOSE: To build and evaluate a small-footprint, lightweight, high-performance 3T MRI scanner for advanced brain imaging with image quality that is equal to or better than conventional whole-body clinical 3T MRI scanners, while achieving substantial reductions in installation costs. METHODS: A conduction-cooled magnet was developed that uses less than 12 liters of liquid helium in a gas-charged sealed system, and standard NbTi wire, and weighs approximately 2000 kg. A 42-cm inner-diameter gradient coil with asymmetric transverse axes was developed to provide patient access for head and extremity exams, while minimizing magnet-gradient interactions that adversely affect image quality. The gradient coil was designed to achieve simultaneous operation of 80-mT/m peak gradient amplitude at a slew rate of 700 T/m/s on each gradient axis using readily available 1-MVA gradient drivers. RESULTS: In a comparison of anatomical imaging in 16 patients using T2 -weighted 3D fluid-attenuated inversion recovery (FLAIR) between the compact 3T and whole-body 3T, image quality was assessed as equivalent to or better across several metrics. The ability to fully use a high slew rate of 700 T/m/s simultaneously with 80-mT/m maximum gradient amplitude resulted in improvements in image quality across EPI, DWI, and anatomical imaging of the brain. CONCLUSIONS: The compact 3T MRI system has been in continuous operation at the Mayo Clinic since March 2016. To date, over 200 patient studies have been completed, including 96 comparison studies with a clinical 3T whole-body MRI. The increased gradient performance has reliably resulted in consistently improved image quality.

Community managed alcohol programs in Canada: Overview of key dimensions and implementation.

INTRODUCTION AND AIMS: People with severe alcohol dependence and unstable housing are vulnerable to multiple harms related to drinking and homelessness. Managed Alcohol Programs (MAP) aim to reduce harms of severe alcohol use without expecting cessation of use. There is promising evidence that MAPs reduce acute and social harms associated with alcohol dependence. The aim of this paper is to describe MAPs in Canada including key dimensions and implementation issues. DESIGN AND METHODS: Thirteen Canadian MAPs were identified through the Canadian Managed Alcohol Program Study. Nine key informant interviews were conducted and analysed alongside program documents and reports to create individual case reports. Inductive content analysis and cross case comparisons were employed to identify six key dimensions of MAPs. RESULTS: Community based MAPs have a common goal of preserving dignity and reducing harms of drinking while increasing access to housing, health and social services. MAPs are offered as both residential and day programs with differences in six key dimensions including program goals and eligibility, food and accomodation, alcohol dispensing and administration, funding and money management, primary care services and clinical monitoring, and social and cultural connections. DISCUSSION AND CONCLUSIONS: MAPs consist of four pillars with the alcohol intervention provided alongside housing interventions, primary care services, social and cultural interventions. Availability of permanent housing and re-establishing social and cultural connections are central to recovery and healing goals of MAPs. Additional research regarding Indigenous and gendered approaches to program development as well as outcomes related to chronic harms and differences in alcohol management are needed.

The effect of concomitant fields in fast spin echo acquisition on asymmetric MRI gradient systems.

PURPOSE: To investigate the effect of the asymmetric gradient concomitant fields (CF) with zeroth and first-order spatial dependence on fast/turbo spin-echo acquisitions, and to demonstrate the effectiveness of their real-time compensation. METHODS: After briefly reviewing the CF produced by asymmetric gradients, the effects of the additional zeroth and first-order CFs on these systems are investigated using extended-phase graph simulations. Phantom and in vivo experiments are performed to corroborate the simulation. Experiments are performed before and after the real-time compensations using frequency tracking and gradient pre-emphasis to demonstrate their effectiveness in correcting the additional CFs. The interaction between the CFs and prescan-based correction to compensate for eddy currents is also investigated. RESULTS: It is demonstrated that, unlike the second-order CFs on conventional gradients, the additional zeroth/first-order CFs on asymmetric gradients cause substantial signal loss and dark banding in fast spin-echo acquisitions within a typical brain-scan field of view. They can confound the prescan correction for eddy currents and degrade image quality. Performing real-time compensation successfully eliminates the artifacts. CONCLUSIONS: We demonstrate that the zeroth/first-order CFs specific to asymmetric gradients can cause substantial artifacts, including signal loss and dark bands for brain imaging. These effects can be corrected using real-time compensation. Magn Reson Med 79:1354-1364, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Nucleophilic (Radio)Fluorination of α-Diazocarbonyl Compounds Enabled by Copper-Catalyzed H-F Insertion.

The copper-catalyzed H-F insertion into α-diazocarbonyl compounds is described using potassium fluoride (KF) and hexafluoroisopropanol. Access to complex α-fluorocarbonyl derivatives is achieved under mild conditions, and the method is readily adapted to radiofluorination with [(18)F]KF. This late-stage strategy provides an attractive route to (18)F-labeled biomolecules.

Conditions that Stabilize Membrane Domains Also Antagonize n-Alcohol Anesthesia.

Diverse molecules induce general anesthesia with potency strongly correlated with both their hydrophobicity and their effects on certain ion channels. We recently observed that several n-alcohol anesthetics inhibit heterogeneity in plasma-membrane-derived vesicles by lowering the critical temperature (Tc) for phase separation. Here, we exploit conditions that stabilize membrane heterogeneity to further test the correlation between the anesthetic potency of n-alcohols and effects on Tc. First, we show that hexadecanol acts oppositely to n-alcohol anesthetics on membrane mixing and antagonizes ethanol-induced anesthesia in a tadpole behavioral assay. Second, we show that two previously described "intoxication reversers" raise Tc and counter ethanol's effects in vesicles, mimicking the findings of previous electrophysiological and behavioral measurements. Third, we find that elevated hydrostatic pressure, long known to reverse anesthesia, also raises Tc in vesicles with a magnitude that counters the effect of butanol at relevant concentrations and pressures. Taken together, these results demonstrate that ΔTc predicts anesthetic potency for n-alcohols better than hydrophobicity in a range of contexts, supporting a mechanistic role for membrane heterogeneity in general anesthesia.