Evaluating Radiation Exposure in Patients with Stable Chest Pain in the SCOT-HEART Trial.

Background In the Scottish Computed Tomography of the Heart (SCOT-HEART) trial in individuals with stable chest pain, a treatment strategy based on coronary CT angiography (CTA) led to improved outcomes. Purpose To assess 5-year cumulative radiation doses of participants undergoing investigation for suspected angina due to coronary artery disease with or without coronary CTA. Materials and Methods This secondary analysis of the SCOT-HEART trial included data from six of 12 recruiting sites and two of three imaging sites. Participants were recruited between November 18, 2010, and September 24, 2014, with follow-up through January 31, 2018. Study participants had been randomized (at a one-to-one ratio) to standard care with CT (n = 1466) or standard care alone (n = 1428). Imaging was performed on a 64-detector (n = 223) or 320-detector row scanner (n = 1466). Radiation dose from CT (dose-length product), SPECT (injected activity), and invasive coronary angiography (ICA; kerma-area product) was assessed for 5 years after enrollment. Effective dose was calculated using conversion factors appropriate for the imaging modality and body region imaged (using 0.026 mSv/mGy · cm for cardiac CT). Results Cumulative radiation dose was assessed in 2894 participants. Median effective dose was 3.0 mSv (IQR, 2.6-3.3 mSv) for coronary calcium scoring, 4.1 mSv (IQR, 2.6-6.1 mSv) for coronary CTA, 7.4 mSv (IQR, 6.2-8.5 mSv) for SPECT, and 4.1 mSv (IQR, 2.5-6.8 mSv) for ICA. After 5 years, total per-participant cumulative dose was higher in the CT group (median, 8.1 mSv; IQR, 5.5-12.4 mSv) compared with standard-care group (median, 0 mSv; IQR, 0-4.5 mSv; P < .001). In participants who underwent any imaging, cumulative radiation exposure was higher in the CT group (n = 1345; median, 8.6 mSv; IQR, 6.1-13.3 mSv) compared with standard-care group (n = 549; median, 6.4 mSv; IQR, 3.4-9.2 mSv; P < .001). Conclusion In the SCOT-HEART trial, the 5-year cumulative radiation dose from cardiac imaging was higher in the coronary CT angiography group compared with the standard-care group, largely because of the radiation exposure from CT. Clinical trial registration no. NCT01149590 © RSNA, 2023 Supplemental material is available for this article. See also the editorial by Dodd and Bosserdt in this issue.

CRISPR screening using an expanded toolkit of autophagy reporters identifies TMEM41B as a novel autophagy factor.

The power of forward genetics in yeast is the foundation on which the field of autophagy research firmly stands. Complementary work on autophagy in higher eukaryotes has revealed both the deep conservation of this process, as well as novel mechanisms by which autophagy is regulated in the context of development, immunity, and neuronal homeostasis. The recent emergence of new clustered regularly interspaced palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-based technologies has begun facilitating efforts to define novel autophagy factors and pathways by forward genetic screening in mammalian cells. Here, we set out to develop an expanded toolkit of autophagy reporters amenable to CRISPR/Cas9 screening. Genome-wide screening of our reporters in mammalian cells recovered virtually all known autophagy-related (ATG) factors as well as previously uncharacterized factors, including vacuolar protein sorting 37 homolog A (VPS37A), transmembrane protein 251 (TMEM251), amyotrophic lateral sclerosis 2 (ALS2), and TMEM41B. To validate this data set, we used quantitative microscopy and biochemical analyses to show that 1 novel hit, TMEM41B, is required for phagophore maturation. TMEM41B is an integral endoplasmic reticulum (ER) membrane protein distantly related to the established autophagy factor vacuole membrane protein 1 (VMP1), and our data show that these two factors play related, albeit not fully overlapping, roles in autophagosome biogenesis. In sum, our work uncovers new ATG factors, reveals a malleable network of autophagy receptor genetic interactions, and provides a valuable resource (http://crispr.deniclab.com) for further mining of novel autophagy mechanisms.

The Get1/2 transmembrane complex is an endoplasmic-reticulum membrane protein insertase.

Hundreds of tail-anchored proteins, including soluble N-ethylmaleimide-sensitive factor attachment receptors (SNAREs) involved in vesicle fusion, are inserted post-translationally into the endoplasmic reticulum membrane by a dedicated protein-targeting pathway. Before insertion, the carboxy-terminal transmembrane domains of tail-anchored proteins are shielded in the cytosol by the conserved targeting factor Get3 (in yeast; TRC40 in mammals). The Get3 endoplasmic-reticulum receptor comprises the cytosolic domains of the Get1/2 (WRB/CAML) transmembrane complex, which interact individually with the targeting factor to drive a conformational change that enables substrate release and, as a consequence, insertion. Because tail-anchored protein insertion is not associated with significant translocation of hydrophilic protein sequences across the membrane, it remains possible that Get1/2 cytosolic domains are sufficient to place Get3 in proximity with the endoplasmic-reticulum lipid bilayer and permit spontaneous insertion to occur. Here we use cell reporters and biochemical reconstitution to define mutations in the Get1/2 transmembrane domain that disrupt tail-anchored protein insertion without interfering with Get1/2 cytosolic domain function. These mutations reveal a novel Get1/2 insertase function, in the absence of which substrates stay bound to Get3 despite their proximity to the lipid bilayer; as a consequence, the notion of spontaneous transmembrane domain insertion is a non sequitur. Instead, the Get1/2 transmembrane domain helps to release substrates from Get3 by capturing their transmembrane domains, and these transmembrane interactions define a bona fide pre-integrated intermediate along a facilitated route for tail-anchor entry into the lipid bilayer. Our work sheds light on the fundamental point of convergence between co-translational and post-translational endoplasmic-reticulum membrane protein targeting and insertion: a mechanism for reducing the ability of a targeting factor to shield its substrates enables substrate handover to a transmembrane-domain-docking site embedded in the endoplasmic-reticulum membrane.

Structural Transformation and Melting in Gold Shock Compressed to 355 GPa.

Gold is believed to retain its ambient crystal structure at very high pressures under static and shock compression, enabling its wide use as a pressure marker. Our in situ x-ray diffraction measurements on shock-compressed gold show that it transforms to the body-centered-cubic (bcc) phase, with an onset pressure between 150 and 176 GPa. A liquid-bcc coexistence was observed between 220 and 302 GPa and complete melting occurs by 355 GPa. Our observation of the lower coordination bcc structure in shocked gold is in marked contrast to theoretical predictions and the reported observation of the hexagonal-close-packed structure under static compression.

Computed tomography myocardial perfusion vs 15O-water positron emission tomography and fractional flow reserve.

OBJECTIVES: Computed tomography (CT) can perform comprehensive cardiac imaging. We compared CT coronary angiography (CTCA) and CT myocardial perfusion (CTP) with 15O-water positron emission tomography (PET) and invasive coronary angiography (ICA) with fractional flow reserve (FFR). METHODS: 51 patients (63 (61-65) years, 80 % male) with known/suspected coronary artery disease (CAD) underwent 320-multidetector CTCA followed by "snapshot" adenosine stress CTP. Of these 22 underwent PET and 47 ICA/FFR. Obstructive CAD was defined as CTCA stenosis >50 % and CTP hypoperfusion, ICA stenosis >70 % or FFR <0.80. RESULTS: PET hyperaemic myocardial blood flow (MBF) was lower in obstructive than non-obstructive territories defined by ICA/FFR (1.76 (1.32-2.20) vs 3.11 (2.44-3.79) mL/(g/min), P < 0.001) and CTCA/CTP (1.76 (1.32-2.20) vs 3.12 (2.44-3.79) mL/(g/min), P < 0.001). Baseline and hyperaemic CT attenuation density was lower in obstructive than non-obstructive territories (73 (71-76) vs 86 (84-88) HU, P < 0.001 and 101 (96-106) vs 111 (107-114) HU, P 0.001). PET hyperaemic MBF corrected for rate pressure product correlated with CT attenuation density (r = 0.579, P < 0.001). There was excellent per-patient sensitivity (96 %), specificity (85 %), negative predictive value (90 %) and positive predictive value (94 %) for CTCA/CTP vs ICA/FFR. CONCLUSION: CT myocardial attenuation density correlates with 15O-water PET MBF. CTCA and CTP can accurately identify obstructive CAD. KEY POINTS: •CT myocardial perfusion can aid the assessment of suspected coronary artery disease. • CT attenuation density from "snapshot" imaging is a marker of myocardial perfusion. • CT myocardial attenuation density correlates with 15 O-water PET myocardial blood flow. • CT attenuation density is lower in obstructive territories defined by invasive angiography. • Diagnostic accuracy of CTCA+CTP is comparable to invasive angiography + fractional flow reserve.

Image quality with single-heartbeat 320-multidetector computed tomographic coronary angiography.

OBJECTIVE: We aimed to establish the feasibility of single-heartbeat 320-multidetector computed tomographic coronary angiography (CTCA) and assess variables affecting image quality. METHODS: Consecutive patients (n = 249, 38% male) underwent CTCA. Two observers assessed image quality using a 4-point scale (1, excellent; 4, poor). RESULTS: Mean heart rate was 60 beats per minute (95% confidence interval, 59-62); body mass index, 29 kg/m (28-30); and dose-length product, 283 mGy·cm (266-301). During scanning, 133 (51%) received sublingual glyceryl trinitrate (GTN), 9 (4%) had ectopics, and 12 (5%) had atrial fibrillation. Diagnostic image quality was obtained in 99% with mean image quality of 1.4 (1.3, 1.5). Age, sex, atrial fibrillation, ectopics, diabetes mellitus (12%), and obstructive disease were not related to image quality. A lower heart rate and GTN were associated with improved image quality (P ≤ 0.001). CONCLUSIONS: Optimal image quality in single-heartbeat 320-multidetector CTCA is achievable in 99% of unselected patients. Image quality is improved by lower heart rate and GTN.

Errors in dual-energy X-ray scanning of the hip because of nonuniform fat distribution.

The variable proportion of fat in overlying soft tissue is a potential source of error in dual-energy X-ray absorptiometry (DXA) measurements of bone mineral. The effect on spine scanning has previously been assessed from cadaver studies and from computed tomography (CT) scans of soft tissue distribution. We have now applied the latter technique to DXA hip scanning. The CT scans performed for clinical purposes were used to derive mean adipose tissue thicknesses over bone and background areas for total hip and femoral neck. The former was always lower. More importantly, the fat thickness differences varied among subjects. Errors because of bone marrow fat were deduced from CT measurements of marrow thickness and assumed fat proportions of marrow. The effect of these differences on measured bone mineral density was deduced from phantom measurements of the bone equivalence of fat. Uncertainties of around 0.06g/cm(2) are similar to those previously reported for spine scanning and the results from cadaver measurements. They should be considered in assessing the diagnostic accuracy of DXA scanning.

Characterization of genes involved in cytokinin signaling and metabolism from rice.

Two-component signaling elements play important roles in plants, including a central role in cytokinin signaling. We characterized two-component elements from the monocot rice (Oryza sativa) using several complementary approaches. Phylogenetic analysis reveals relatively simple orthologous relationships among the histidine kinases in rice and Arabidopsis (Arabidopsis thaliana). In contrast, the histidine-containing phosphotransfer proteins (OsHPs) and response regulators (OsRRs) display a higher degree of lineage-specific expansion. The intracellular localizations of several OsHPs and OsRRs were examined in rice and generally found to correspond to the localizations of their dicot counterparts. The functionality of rice type-B OsRRs was tested in Arabidopsis; one from a clade composed of both monocot and dicot type-B OsRRs complemented an Arabidopsis type-B response regulator mutant, but a type-B OsRR from a monocot-specific subfamily generally did not. The expression of genes encoding two-component elements and proteins involved in cytokinin biosynthesis and degradation was analyzed in rice roots and shoots and in response to phytohormones. Nearly all type-A OsRRs and OsHK4 were up-regulated in response to cytokinin, but other cytokinin signaling elements were not appreciably affected. Furthermore, multiple cytokinin oxidase (OsCKX) genes were up-regulated by cytokinin. Abscisic acid treatment decreased the expression of several genes involved in cytokinin biosynthesis and degradation. Auxin affected the expression of a few genes; brassinosteroid and gibberellin had only modest effects. Our results support a shared role for two-component elements in mediating cytokinin signaling in monocots and dicots and reveal how phytohormones can impact cytokinin function through modulating gene expression.

A field programmable gate array-based timing and control system for the dynamic compression sector.

A field programmable gate array (FPGA)-based timing and trigger control system has been developed for the Dynamic Compression Sector (DCS) user facility located at the Advanced Photon Source (APS) at Argonne National Laboratory. The DCS is a first-of-its-kind capability dedicated to dynamic compression science. All components of the DCS laser shock station-x-ray choppers, single-shot shutter, internal laser triggers, and shot diagnostics-must be synchronized with respect to the arrival of x rays in the hutch. An FPGA synchronized to the APS storage ring radio frequency clock (352 MHz) generates trigger signals for each stage of the laser and x-ray shutter system with low jitter. The developed FPGA-based control system was the first system used to control the laser and the shutter system since its commissioning, and it has been developing since then to improve the timing jitter. The system is composed of a Zynq FPGA, a debug card, line drivers, and a power supply. The delay and offsets of the trigger signals can be adjusted by using a user-friendly graphical user interface with high precision. The details of the system architecture, timing requirements, firmware, and software implementation along with the performance evaluation are presented in this paper. The system offers low timing jitter (15.5 ps rms) with respect to the APS 352 MHz clock, suitable for the 100 ps (FWHM) x-ray bunch duration at the APS.

The laser shock station in the dynamic compression sector. I.

The Laser Shock Station in the Dynamic Compression Sector (DCS) [Advanced Photon Source (APS), Argonne National Laboratory] links a laser-driven shock compression platform with high energy x-ray pulses from the APS to achieve in situ, time-resolved x-ray measurements (diffraction and imaging) in materials subjected to well-characterized, high stress, short duration shock waves. This station and the other DCS experimental stations provide a unique and versatile facility to study condensed state phenomena subjected to shocks with a wide range of amplitudes (to above ∼350 GPa) and time-durations (∼10 ns-1 µs). The Laser Shock Station uses a 100 J, 5-17 ns, 351 nm frequency tripled Nd:glass laser with programmable pulse shaping and focal profile smoothing for maximum precision. The laser can operate once every 30 min. The interaction chamber has multiple diagnostic ports, a sample holder to expose 14 samples without breaking vacuum, can vary the angle between the x-ray and laser beams by 135°, and can translate to select one of the two types of x-ray beams. The x-ray measurement temporal resolution is ∼90 ps. The system is capable of reproducible, well-characterized experiments. In a series of 10 shots, the absolute variation in shock breakout times was less than 500 ps. The variation in peak particle velocity at the sample/window interface was 4.3%. This paper describes the entire DCS Laser Shock Station, including sample fabrication and diagnostics, as well as experimental results from shock compressed tantalum that demonstrate the facility's capability for acquiring high quality x-ray diffraction data.

Retro-2 protects cells from ricin toxicity by inhibiting ASNA1-mediated ER targeting and insertion of tail-anchored proteins.

The small molecule Retro-2 prevents ricin toxicity through a poorly-defined mechanism of action (MOA), which involves halting retrograde vesicle transport to the endoplasmic reticulum (ER). CRISPRi genetic interaction analysis revealed Retro-2 activity resembles disruption of the transmembrane domain recognition complex (TRC) pathway, which mediates post-translational ER-targeting and insertion of tail-anchored (TA) proteins, including SNAREs required for retrograde transport. Cell-based and in vitro assays show that Retro-2 blocks delivery of newly-synthesized TA-proteins to the ER-targeting factor ASNA1 (TRC40). An ASNA1 point mutant identified using CRISPR-mediated mutagenesis abolishes both the cytoprotective effect of Retro-2 against ricin and its inhibitory effect on ASNA1-mediated ER-targeting. Together, our work explains how Retro-2 prevents retrograde trafficking of toxins by inhibiting TA-protein targeting, describes a general CRISPR strategy for predicting the MOA of small molecules, and paves the way for drugging the TRC pathway to treat broad classes of viruses known to be inhibited by Retro-2.

The AAA protein Msp1 mediates clearance of excess tail-anchored proteins from the peroxisomal membrane.

Msp1 is a conserved AAA ATPase in budding yeast localized to mitochondria where it prevents accumulation of mistargeted tail-anchored (TA) proteins, including the peroxisomal TA protein Pex15. Msp1 also resides on peroxisomes but it remains unknown how native TA proteins on mitochondria and peroxisomes evade Msp1 surveillance. We used live-cell quantitative cell microscopy tools and drug-inducible gene expression to dissect Msp1 function. We found that a small fraction of peroxisomal Pex15, exaggerated by overexpression, is turned over by Msp1. Kinetic measurements guided by theoretical modeling revealed that Pex15 molecules at mitochondria display age-independent Msp1 sensitivity. By contrast, Pex15 molecules at peroxisomes are rapidly converted from an initial Msp1-sensitive to an Msp1-resistant state. Lastly, we show that Pex15 interacts with the peroxisomal membrane protein Pex3, which shields Pex15 from Msp1-dependent turnover. In sum, our work argues that Msp1 selects its substrates on the basis of their solitary membrane existence.

Iterative reconstruction can permit the use of lower X-ray tube current in CT coronary artery calcium scoring.

OBJECTIVE:: CT coronary artery calcium scoring (CACS) is additive to traditional risk factors for predicting future cardiac events but is associated with relatively high radiation doses. We assessed the feasibility of CACS radiation dose reduction using a lower tube current and iterative reconstruction (IR). METHODS:: Artificial noise was added to the raw data from 27 CACS studies from patients who were symptomatic to simulate lower tube current scanning (75, 50 and 25% original current). All studies were performed on the same CT scanner at 120 kVp. Data were reconstructed using filtered back projection [Quantum Denoising Software (QDS+)] and IR [adaptive iterative dose reduction three dimensional mild, standard and strong]. Agatston scores were independently measured by two readers. CACS percentile risk scores were calculated. RESULTS:: At 75, 50 and 25% tube currents, all adaptive iterative dose reduction (AIDR) reconstructions decreased image noise relative to QDS+ (p < 0.05). All AIDR reconstructions resulted in small reductions in Agatston score relative to QDS+ at the standard tube current (p < 0.05). Agatston scores increased with QDS+ at 75, 50 and 25% tube current (p < 0.05), whereas no significant change was observed with AIDR mild at any tested tube current. No difference in the percentile risk score with AIDR mild at any tube current occurred compared with QDS+ at standard tube current (p > 0.05). Interobserver agreement for AIDR mild remained excellent even at 25% tube current (intraclass correlation coefficient 0.997). CONCLUSION:: Up to 75% reduction in CACS tube current is feasible using AIDR mild. ADVANCES IN KNOWLEDGE:: AIDR mild IR permits low tube current CACS whilst maintaining excellent intraobserver and interobserver variability and without altering risk classification.

AngioCT in the management of neurointerventional patients: a prospective, consecutive series with associated dosimetry and resolution data.

INTRODUCTION: Endovascular coiling of intracranial aneurysms carries a risk of complications. Early detection and management of complications can improve clinical outcomes. AngioCT is a new imaging technology enabling CT-like images to be generated on a flat-panel digital subtraction angiography system, which can provide immediate "on angio table" identification and thorough assessment of such complications. We prospectively audited its utility during aneurysm coiling in patients following subarachnoid haemorrhage (SAH). METHODS: A prospective series of 44 patients with SAH undergoing endovascular coiling with AngioCT was audited for image quality and the influence of the AngioCT on patient management. In a parallel experimental study, radiation doses were measured and image quality parameters on standard phantoms were established. RESULTS: In all patients, AngioCT provided adequate diagnostic information. In 40.9% of patients, AngioCT was a substantial or major factor in determining the management immediately after coiling. Using a 10-s high-dose acquisition technique, acceptable image quality could be obtained rapidly with a radiation dose just over half that for a conventional CT scan of the head (35 mGy versus approximately 60 mGy). No patient in this series required conventional CT to clarify the AngioCT appearance. CONCLUSION: AngioCT has many applications in the neurointerventional setting. In particular during coiling, AngioCT provides a rapid way to clarify concerns or identify complications and in some cases was the major factor influencing further patient management immediately after coiling. AngioCT images were judged of adequate quality to be clinically useful in all patients in this series.