Optimization of upstream particle concentration from flow using AC electro-osmosis and dielectrophoresis.

There are many applications where upstream sample processing is required to concentrate dispersed particles in flow; this may be to increase the concentration (e.g., to enhance biosensor accuracy) or to decrease it (e.g., by removing contaminants from flow). The AC electrokinetic phenomenon, dielectrophoresis (DEP), has been used widely for particle trapping for flow, but the magnitude of the force drops reduces rapidly with distance from electrode edges, so that nm-scale particles such as viruses and bacteria are only trapped when near the electrode surface. This limits the usable flow rate in the device and can render the final device unusable for practical applications. Conversely, another electrokinetic phenomenon, AC electro-osmosis (ACEO), can be used to move particles to electrode surfaces but is unable to trap them from flow, limiting their ability for sample cleanup or trap-and-purge concentration. In this paper, we describe the optimization of ACEO electrodes aligned parallel to pressure-driven flow as a precursor/preconditioner to capture particles from a flow stream and concentrate them adjacent to the channel wall to enhance DEP capture. This is shown to be effective at flow rates of up to 0.84 ml min-1. Furthermore, the analysis of the 3D flow structure in the ACEO device by both simulation and confocal microscopy suggests that while the system offers significant benefits, the flow structure in the volume near the channel lid is such that while substantial trapping can occur, particles in this part of the chamber cannot be trapped, independent of the chamber height.

Photosynthetic control at the cytochrome b6f complex.

Photosynthetic control (PCON) is a protective mechanism that prevents light-induced damage to photosystem I (PSI) by ensuring the rate of NADPH and ATP production via linear electron transfer (LET) is balanced by their consumption in the CO2 fixation reactions. Protection of PSI is a priority for plants since they lack a dedicated rapid-repair cycle for this complex, meaning that any damage leads to prolonged photoinhibition and decreased growth. The imbalance between LET and the CO2 fixation reactions is sensed at the level of the transthylakoid ΔpH, which increases when light is in excess. The canonical mechanism of PCON involves feedback control by ΔpH on the plastoquinol oxidation step of LET at cytochrome b6f. PCON thereby maintains the PSI special pair chlorophylls (P700) in an oxidized state, that allows excess electrons unused in the CO2 fixation reactions to be safely quenched via charge recombination. In this review we focus on angiosperms, considering how photo-oxidative damage to PSI comes about, explore the consequences of PSI photoinhibition on photosynthesis and growth, discuss recent progress in understanding PCON regulation, and finally consider the prospects for its future manipulation in crop plants to improve photosynthetic efficiency.

Dielectric properties of human macrophages are altered by Mycobacterium tuberculosis infection.

The analysis of cell electrophysiology for pathogenic samples at BSL3 can be problematic. It is virtually impossible to isolate infected from uninfected without a label, for example green fluorescent protein, which can potentially alter the cell electrical properties. Furthermore, the measurement of highly pathogenic organisms often requires equipment dedicated only for use with these organisms due to safety considerations. To address this, we have used dielectrophoresis to study the electrical properties of the human THP-1 cell line and monocyte-derived macrophages before and after infection with non-labelled Mycobacterium tuberculosis. Infection with these highly pathogenic bacilli resulted in changes including a raised surface conductance (associated with reduced zeta potential) and increased capacitance, suggesting an increase in surface roughness. We have also investigated the effect of fixation on THP-1 cells as a means to enable study on fixed samples in BSL1 or 2 laboratories, which suggests that the properties of these cells are largely unaffected by the fixation process. This advance results in a novel technique enabling the isolation of infected and non-infected cells in a sample without labelling.

Targeted Training Reduces Search Errors but Not Classification Errors for Hepatic Metastasis Detection at Contrast-Enhanced CT.

RATIONALE AND OBJECTIVES: Methods are needed to improve the detection of hepatic metastases. Errors occur in both lesion detection (search) and decisions of benign versus malignant (classification). Our purpose was to evaluate a training program to reduce search errors and classification errors in the detection of hepatic metastases in contrast-enhanced abdominal computed tomography (CT). MATERIALS AND METHODS: After Institutional Review Board approval, we conducted a single-group prospective pretest-posttest study. Pretest and posttest were identical and consisted of interpreting 40 contrast-enhanced abdominal CT exams containing 91 liver metastases under eye tracking. Between pretest and posttest, readers completed search training with eye-tracker feedback and coaching to increase interpretation time, use liver windows, and use coronal reformations. They also completed classification training with part-task practice, rating lesions as benign or malignant. The primary outcome was metastases missed due to search errors (<2 seconds gaze under eye tracker) and classification errors (>2 seconds). Jackknife free-response receiver operator characteristic (JAFROC) analysis was also conducted. RESULTS: A total of 31 radiologist readers (8 abdominal subspecialists, 8 nonabdominal subspecialists, 15 senior residents/fellows) participated. Search errors were reduced (pretest 11%, posttest 8%, difference 3% [95% confidence interval, 0.3%-5.1%], P = .01), but there was no difference in classification errors (difference 0%, P = .97) or in JAFROC figure of merit (difference -0.01, P = .36). In subgroup analysis, abdominal subspecialists demonstrated no evidence of change. CONCLUSION: Targeted training reduced search errors but not classification errors for the detection of hepatic metastases at contrast-enhanced abdominal CT. Improvements were not seen in all subgroups.

Proton Gradient Regulation 5 is required to avoid photosynthetic oscillations during light transitions.

The production of ATP and NADPH by the light reactions of photosynthesis and their consumption by the Calvin-Benson-Bassham (CBB) cycle and other downstream metabolic reactions requires careful regulation. Environmental shifts perturb this balance, leading to photo-oxidative stress and losses in CO2 assimilation. Imbalances in the production and consumption of ATP and NADPH manifest themselves as transient instability in the chlorophyll fluorescence, P700, electrochromic shift, and CO2 uptake signals recorded on leaves. These oscillations can be induced in wild-type plants by sudden shifts in CO2 concentration or light intensity; however, mutants exhibiting increased oscillatory behaviour have yet to be reported. This has precluded an understanding of the regulatory mechanisms employed by plants to suppress oscillations. Here we show that the Arabidopsis pgr5 mutant, which is deficient in Proton Gradient Regulation 5 (PGR5)-dependent cyclic electron transfer (CET), exhibits increased oscillatory behaviour. In contrast, mutants lacking the NADH-dehydrogenase-like-dependent CET are largely unaffected. The absence of oscillations in the hope2 mutant which, like pgr5, lacks photosynthetic control and exhibits high ATP synthase conductivity, ruled out loss of these photoprotective mechanisms as causes. Instead, we observed slower formation of the proton motive force and, by inference, ATP synthesis in pgr5 following environmental perturbation, leading to the transient reduction of the electron transfer chain and photosynthetic oscillations. PGR5-dependent CET therefore plays a major role in damping the effect of environmental perturbations on photosynthesis to avoid losses in CO2 fixation.

Infrapopliteal Segments on Lower Extremity CTA: Prospective Intraindividual Comparison of Energy-Integrating Detector CT and Photon-Counting Detector CT.

BACKGROUND. The higher spatial resolution and image contrast for iodine-containing tissues of photon-counting detector (PCD) CT may address challenges in evaluating small calcified vessels when performing lower extremity CTA by energy-integrating detector (EID) CTA. OBJECTIVE. The purpose of the study was to compare the evaluation of infrapopliteal vasculature between lower extremity CTA performed using EID CT and PCD CT. METHODS. This prospective study included 32 patients (mean age, 69.7 ± 11.3 [SD] years; 27 men, five women) who underwent clinically indicated lower extremity EID CTA between April 2021 and March 2022; participants underwent investigational lower extremity PCD CTA later the same day as EID CTA using a reduced IV contrast media dose. Two radiologists independently reviewed examinations in two sessions, each containing a random combination of EID CTA and PCD CTA examinations; the readers assessed the number of visualized fibular perforators, characteristics of stenoses at 11 infrapopliteal segmental levels, and subjective arterial sharpness. RESULTS. Mean IV contrast media dose was 60.0 ± 11.0 (SD) mL for PCD CTA versus 139.6 ± 11.8 mL for EID CTA (p < .001). The number of identified fibular perforators per lower extremity was significantly higher for PCD CTA than for EID CTA for reader 1 (R1) (mean ± SD, 6.4 ± 3.2 vs 4.2 ± 2.4; p < .001) and reader 2 (R2) (8.8 ± 3.4 vs 7.6 ± 3.3; p = .04). Reader confidence for assessing stenosis was significantly higher for PCD CTA than for EID CTA for R1 (mean ± SD, 82.3 ± 20.3 vs 78.0 ± 20.2; p < .001) but not R2 (89.8 ± 16.7 vs 90.6 ± 7.1; p = .24). The number of segments per lower extremity with total occlusion was significantly lower for PCD CTA than for EID CTA for R2 (mean ± SD, 0.5 ± 1.3 vs 0.9 ± 1.7; p = .04) but not R1 (0.6 ± 1.3 vs 1.0 ± 1.5; p = .07). The number of segments per lower extremity with clinically significant nonocclusive stenosis was significantly higher for PCD CTA than for EID CTA for R1 (mean ± SD, 2.2 ± 2.2 vs 1.6 ± 1.7; p = .01) but not R2 (1.1 ± 2.0 vs 1.1 ± 1.4; p = .89). Arterial sharpness was significantly greater for PCD CTA than for EID CTA for R1 (mean ± SD, 3.2 ± 0.5 vs 1.8 ± 0.5; p < .001) and R2 (3.2 ± 0.4 vs 1.7 ± 0.8; p < .001). CONCLUSION. PCD CTA yielded multiple advantages relative to EID CTA for visualizing small infrapopliteal vessels and characterizing associated plaque. CLINICAL IMPACT. The use of PCD CTA may improve vascular evaluation in patients with peripheral arterial disease.

Energetic driving force for LHCII clustering in plant membranes.

Plants capture and convert solar energy in a complex network of membrane proteins. Under high light, the luminal pH drops and induces a reorganization of the protein network, particularly clustering of the major light-harvesting complex (LHCII). While the structures of the network have been resolved in exquisite detail, the thermodynamics that control the assembly and reorganization had not been determined, largely because the interaction energies of membrane proteins have been inaccessible. Here, we describe a method to quantify these energies and its application to LHCII. Using single-molecule measurements, LHCII proteoliposomes, and statistical thermodynamic modeling, we quantified the LHCII-LHCII interaction energy as ~-5 kBT at neutral pH and at least -7 kBT at acidic pH. These values revealed an enthalpic thermodynamic driving force behind LHCII clustering. Collectively, this work captures the interactions that drive the organization of membrane protein networks from the perspective of equilibrium statistical thermodynamics, which has a long and rich tradition in biology.

Incidence of complications related to shoulder arthroplasties identified on computed tomography.

PURPOSE: Determine incidence of shoulder arthroplasty complications identified on computed tomography (CT). MATERIALS AND METHODS: Retrospective institutional database review of patients with shoulder arthroplasties who underwent CT between 01/2006-11/2021 at a tertiary academic referral center with subspecialized orthopedic shoulder surgeons. CT reports were reviewed for arthroplasty type and complication. Data were stratified and summarized. Associations between complications and arthroplasty types were determined with Chi-squared goodness of fit test. RESULTS: Eight hundred twelve CTs in 797 unique patients were included (438 (53.9%) females and 374 (46.1%) males; mean age 67 ± 11 years). There were 403 total shoulder arthroplasties (TSA), 317 reverse total shoulder arthroplasties (rTSA), and 92 hemiarthroplasties (HA). Complications were present in 527/812 (64.9%) and incidences were: loosening/aseptic osteolysis 36.9%, periprosthetic failure 21.6%, periprosthetic fracture 12.3%, periprosthetic dislocation 6.8%, joint/pseudocapsule effusion 5.9%, prosthetic failure 4.8%, infection 3.8%, and periprosthetic collection 2.1%. Complications per arthroplasty were: 305/403 (75.7%) TSAs, 176/317 (55.5%) rTSAs, and 46/92 (50%) HAs (p < 0.001). Periprosthetic fracture (20.8%), prosthetic dislocation (9.8%), and prosthetic failure (7.9%) were highest in rTSAs (p < 0.001, p < 0.013, p < 0.001, respectively). Loosening/aseptic osteolysis most frequent in TSAs (54.1%) (p < 0.001). Periprosthetic failure most frequent in HA (32.6%) (p < 0.001). Significant associations were identified with joint/pseudocapsule effusion and loosening/aseptic osteolysis (p = 0.04) and prosthetic dislocation (p < .001). CONCLUSION: In this single tertiary academic referral center cohort, the incidence of shoulder arthroplasty complication identified on CT was 64.9% and the most commonly occurring complication was loosening/aseptic osteolysis (36.9%). TSA had the highest incidence of complication (75.7%).

Twisted Carotenoids Do Not Support Efficient Intramolecular Singlet Fission in the Orange Carotenoid Protein.

Singlet exciton fission is the spin-allowed generation of two triplet electronic excited states from a singlet state. Intramolecular singlet fission has been suggested to occur on individual carotenoid molecules within protein complexes provided that the conjugated backbone is twisted out of plane. However, this hypothesis has been forwarded only in protein complexes containing multiple carotenoids and bacteriochlorophylls in close contact. To test the hypothesis on twisted carotenoids in a "minimal" one-carotenoid system, we study the orange carotenoid protein (OCP). OCP exists in two forms: in its orange form (OCPo), the single bound carotenoid is twisted, whereas in its red form (OCPr), the carotenoid is planar. To enable room-temperature spectroscopy on canthaxanthin-binding OCPo and OCPr without laser-induced photoconversion, we trap them in a trehalose glass. Using transient absorption spectroscopy, we show that there is no evidence of long-lived triplet generation through intramolecular singlet fission despite the canthaxanthin twist in OCPo.

Outcomes in patients with clinically suspected pedal osteomyelitis based on bone marrow signal pattern on MRI.

Objective: confluent T1 hypointense marrow signal is widely accepted to represent osteomyelitis on MRI. Some authors have suggested that non-confluent bone marrow signal abnormality should be considered early osteomyelitis. The purpose of this study was to address this issue by comparing the rate of osteomyelitis and amputation based on T1 marrow signal characteristics. Materials and methods: a total of 112 patients who underwent MRI of the foot for the evaluation of possible osteomyelitis were included. Patients were assigned to confluent T1 hypointense, reticulated T1 hypointense, and normal bone marrow signal groups. Results: patients with confluent T1 hypointense signal on MRI had significantly higher rates of osteomyelitis and amputation at 2 and 14 months post-MRI than the reticulated T1 hypointense group ( p < 0.001 ). Six patients had normal T1 signal, 16.7 % of whom had osteomyelitis and underwent amputation by 2 months post-MRI. Of 61 patients with reticulated T1 hypointense signal, 19.7 % had a diagnosis of osteomyelitis at 2 months post-MRI and 30.8 % had a diagnosis of osteomyelitis at 14 months post-MRI; moreover, 14.8 % and 31.5 % underwent amputation by 2 and 14 months post-MRI, respectively. Of 45 patients with confluent T1 hypointense signal, 73.3 % of patients had osteomyelitis at 2 months post-MRI and 82.5 % had osteomyelitis at 14 months post-MRI. In this group, 66.7 % underwent amputation by 2 months post-MRI and 77.8 % underwent amputation by 14 months post-MRI. Conclusions: over half of the patients with suspected pedal osteomyelitis who had reticulated or normal T1 bone marrow signal on MRI healed with conservative measures. Therefore, we recommend terminology such as "osteitis", "reactive osteitis", or "nonspecific reactive change" to describe bone marrow edema-like signal and reticulated hazy T1 hypointense signal without associated confluent T1 hypointensity. Moreover, we recommend that the MRI diagnosis of osteomyelitis is reserved for confluent T1 hypointense bone signal in the area of concern.

A training program to reduce reader search errors for liver metastasis detection in CT.

Detection of low contrast liver metastases varies between radiologists. Training may improve performance for lower-performing readers and reduce inter-radiologist variability. We recruited 31 radiologists (15 trainees, 8 non-abdominal staff, and 8 abdominal staff) to participate in four separate reading sessions: pre-test, search training, classification training, and post-test. In the pre-test, each radiologist interpreted 40 liver CT exams containing 91 metastases, circumscribed suspected hepatic metastases while under eye tracker observation, and rated confidence. In search training, radiologists interpreted a separate set of 30 liver CT exams while receiving eye tracker feedback and after coaching to increase use of coronal reformations, interpretation time, and use of liver windows. In classification training, radiologists interpreted up to 100 liver CT image patches, most with benign or malignant lesions, and compared their annotations to ground truth. Post-test was identical to pre-test. Between pre- and post-test, sensitivity increased by 2.8% (p = 0.01) but AUC did not change significantly. Missed metastases were classified as search errors (<2 seconds gaze time) or classification errors (>2 seconds gaze time) using the eye tracker. Out of 2775 possible detections, search errors decreased (10.8% to 8.1%; p < 0.01) but classification errors were unchanged (5.7% vs 5.7%). When stratified by difficulty, easier metastases showed larger reductions in search errors: for metastases with average sensitivity of 0-50%, 50-90%, and 90-100%, reductions in search errors were 16%, 35%, and 58%, respectively. The training program studied here may be able to improve radiologist performance by reducing errors but not classification errors.

STN7 is not essential for developmental acclimation of Arabidopsis to light intensity.

Plants respond to changing light intensity in the short term through regulation of light harvesting, electron transfer, and metabolism to mitigate redox stress. A sustained shift in light intensity leads to a long-term acclimation response (LTR). This involves adjustment in the stoichiometry of photosynthetic complexes through de novo synthesis and degradation of specific proteins associated with the thylakoid membrane. The light-harvesting complex II (LHCII) serine/threonine kinase STN7 plays a key role in short-term light harvesting regulation and was also suggested to be crucial to the LTR. Arabidopsis plants lacking STN7 (stn7) shifted to low light experience higher photosystem II (PSII) redox pressure than the wild type or those lacking the cognate phosphatase TAP38 (tap38), while the reverse is true at high light, where tap38 suffers more. In principle, the LTR should allow optimisation of the stoichiometry of photosynthetic complexes to mitigate these effects. We used quantitative label-free proteomics to assess how the relative abundance of photosynthetic proteins varied with growth light intensity in wild-type, stn7, and tap38 plants. All plants were able to adjust photosystem I, LHCII, cytochrome b6 f, and ATP synthase abundance with changing white light intensity, demonstrating neither STN7 nor TAP38 is crucial to the LTR per se. However, stn7 plants grown for several weeks at low light (LL) or moderate light (ML) still showed high PSII redox pressure and correspondingly lower PSII efficiency, CO2 assimilation, and leaf area compared to wild-type and tap38 plants, hence the LTR is unable to fully ameliorate these symptoms. In contrast, under high light growth conditions the mutants and wild type behaved similarly. These data are consistent with the paramount role of STN7-dependent LHCII phosphorylation in tuning PSII redox state for optimal growth in LL and ML conditions.

Cryo-EM structure of the four-subunit Rhodobacter sphaeroides cytochrome bc1 complex in styrene maleic acid nanodiscs.

Cytochrome bc1 complexes are ubiquinol:cytochrome c oxidoreductases, and as such, they are centrally important components of respiratory and photosynthetic electron transfer chains in many species of bacteria and in mitochondria. The minimal complex has three catalytic components, which are cytochrome b, cytochrome c1, and the Rieske iron-sulfur subunit, but the function of mitochondrial cytochrome bc1 complexes is modified by up to eight supernumerary subunits. The cytochrome bc1 complex from the purple phototrophic bacterium Rhodobacter sphaeroides has a single supernumerary subunit called subunit IV, which is absent from current structures of the complex. In this work we use the styrene-maleic acid copolymer to purify the R. sphaeroides cytochrome bc1 complex in native lipid nanodiscs, which retains the labile subunit IV, annular lipids, and natively bound quinones. The catalytic activity of the four-subunit cytochrome bc1 complex is threefold higher than that of the complex lacking subunit IV. To understand the role of subunit IV, we determined the structure of the four-subunit complex at 2.9 Å using single particle cryogenic electron microscopy. The structure shows the position of the transmembrane domain of subunit IV, which lies across the transmembrane helices of the Rieske and cytochrome c1 subunits. We observe a quinone at the Qo quinone-binding site and show that occupancy of this site is linked to conformational changes in the Rieske head domain during catalysis. Twelve lipids were structurally resolved, making contacts with the Rieske and cytochrome b subunits, with some spanning both of the two monomers that make up the dimeric complex.

High cyclic electron transfer via the PGR5 pathway in the absence of photosynthetic control.

The light reactions of photosynthesis couple electron and proton transfers across the thylakoid membrane, generating NADPH, and proton motive force (pmf) that powers the endergonic synthesis of ATP by ATP synthase. ATP and NADPH are required for CO2 fixation into carbohydrates by the Calvin-Benson-Bassham cycle. The dominant ΔpH component of the pmf also plays a photoprotective role in regulating photosystem II light harvesting efficiency through nonphotochemical quenching (NPQ) and photosynthetic control via electron transfer from cytochrome b6f (cytb6f) to photosystem I. ΔpH can be adjusted by increasing the proton influx into the thylakoid lumen via upregulation of cyclic electron transfer (CET) or decreasing proton efflux via downregulation of ATP synthase conductivity (gH+). The interplay and relative contributions of these two elements of ΔpH control to photoprotection are not well understood. Here, we showed that an Arabidopsis (Arabidopsis thaliana) ATP synthase mutant hunger for oxygen in photosynthetic transfer reaction 2 (hope2) with 40% higher proton efflux has supercharged CET. Double crosses of hope2 with the CET-deficient proton gradient regulation 5 and ndh-like photosynthetic complex I lines revealed that PROTON GRADIENT REGULATION 5 (PGR5)-dependent CET is the major pathway contributing to higher proton influx. PGR5-dependent CET allowed hope2 to maintain wild-type levels of ΔpH, CO2 fixation and NPQ, however photosynthetic control remained absent and PSI was prone to photoinhibition. Therefore, high CET in the absence of ATP synthase regulation is insufficient for PSI photoprotection.

Evaluating Variability of Commercial Liver Fibrosis Elastography Phantoms.

OBJECTIVE: Liver fibrosis has been found to increase the mechanical stiffness of the liver. To mimic different stages of liver fibrosis, commercially available phantoms (Model 039, CIRS, Inc.) have been produced for clinical quality assurance and research purposes. The purpose of this study was to investigate the mechanical property variability of the phantoms in two lots of CIRS Model 039 phantoms. METHODS: Each lot consisted of phantoms of four stiffness types, and there were 8-10 phantoms of each type. Shear wave elastography measurements were conducted on each phantom at 10 different angles. Group velocity measurements and phase velocity curves were calculated for every SWE acquisition. Multilevel functional principal component analysis (MFPCA) was performed on phase velocity data, which decomposes each phase velocity curve into the sum of eigenfunctions of two levels. The variance of the component scores of levels 1 and 2 were used to represent inter-phantom and intra-phantom variability, respectively. The 95% confidence intervals of phase velocity in a phantom type were calculated to reflect curve variability. DISCUSSION: The standard deviations of the group velocity for phantoms of any type were less than 0.04 and 0.02 m/s for lots 1 and 2, respectively. For both lots, in every type, the phase velocity curves of most individual phantoms fall within the 95% confidence interval. CONCLUSION: MFPCA is an effective tool for analyzing the inter- and intra-phantom variability of phase velocity curves. Given the known variability of a fully tested lot, estimation of the variability of a new lot can be performed with a reduced number of phantoms tested.

Diagnostic Performance in Low- and High-Contrast Tasks of an Image-Based Denoising Algorithm Applied to Radiation Dose-Reduced Multiphase Abdominal CT Examinations.

BACKGROUND. Anatomic redundancy between phases can be used to achieve denoising of multiphase CT examinations. A limitation of iterative reconstruction (IR) techniques is that they generally require use of CT projection data. A frequency-split multi-band-filtration algorithm applies denoising to the multiphase CT images themselves. This method does not require knowledge of the acquisition process or integration into the reconstruction system of the scanner, and it can be implemented as a supplement to commercially available IR algorithms. OBJECTIVE. The purpose of the present study is to compare radiologists' performance for low-contrast and high-contrast diagnostic tasks (i.e., tasks for which differences in CT attenuation between the imaging target and its anatomic background are subtle or large, respectively) evaluated on multiphase abdominal CT between routine-dose images and radiation dose-reduced images processed by a frequency-split multiband-filtration denoising algorithm. METHODS. This retrospective single-center study included 47 patients who underwent multiphase contrast-enhanced CT for known or suspected liver metastases (a low-contrast task) and 45 patients who underwent multiphase contrast-enhanced CT for pancreatic cancer staging (a high-contrast task). Radiation dose-reduced images corresponding to dose reduction of 50% or more were created using a validated noise insertion technique and then underwent denoising using the frequency-split multi-band-filtration algorithm. Images were independently evaluated in multiple sessions by different groups of abdominal radiologists for each task (three readers in the low-contrast arm and four readers in the high-contrast arm). The noninferiority of denoised radiation dose-reduced images to routine-dose images was assessed using the jackknife alternative free-response ROC (JAFROC) figure-of-merit (FOM; limit of noninferiority, -0.10) for liver metastases detection and using the Cohen kappa statistic and reader confidence scores (100-point scale) for pancreatic cancer vascular invasion. RESULTS. For liver metastases detection, the JAFROC FOM for denoised radiation dose-reduced images was 0.644 (95% CI, 0.510-0.778), and that for routine-dose images was 0.668 (95% CI, 0.543-0.792; estimated difference, -0.024 [95% CI, -0.084 to 0.037]). Intraobserver agreement for pancreatic cancer vascular invasion was substantial to near perfect when the two image sets were compared (κ = 0.53-1.00); the 95% CIs of all differences in confidence scores between image sets contained zero. CONCLUSION. Multiphase contrast-enhanced abdominal CT images with a radiation dose reduction of 50% or greater that undergo denoising by a frequency-split multiband-filtration algorithm yield performance similar to that of routine-dose images for detection of liver metastases and vascular staging of pancreatic cancer. CLINICAL IMPACT. The image-based denoising algorithm facilitates radiation dose reduction of multiphase examinations for both low- and high-contrast diagnostic tasks without requiring manufacturer-specific hardware or software.

PPV of the Molecular Breast Imaging Lexicon.

BACKGROUND. Molecular breast imaging (MBI) is used for various breast imaging indications. An MBI lexicon has been developed, although the likelihood of malignancy of the lexicon descriptors has not been assessed to our knowledge. OBJECTIVE. The purpose of this article was to evaluate the PPV for malignancy of the MBI lexicon imaging descriptors. METHODS. This retrospective study included MBI examinations performed from August 1, 2005, through August 31, 2017, that were positive (BI-RADS analogous categories 0, 3, 4, 5, or 6) according to the clinical report and had an available reference standard. Examinations were performed using dual-detector cadmium zinc telluride MBI systems after injection of 99mTc sestamibi. Category 3 lesions had pathologic correlation, at least 2 years of imaging follow-up, or final resolution on follow-up imaging as category 1 or 2; category 4 and 5 lesions had pathologic correlation. MBI examinations were reviewed by one of two radiologists to assess lesions on the basis of the published MBI lexicon for type (mass vs nonmass uptake), distribution (if nonmass uptake), uptake intensity, and number of MBI views on which the lesion was seen. PPV for malignancy was summarized. RESULTS. The analysis included 643 lesions (479 benign, 164 malignant; 83 mass, 560 nonmass uptake) in 509 patients (median age, 56 years). PPV was 73.5% (61/83) for masses and 18.4% (103/560) for nonmass uptake. Among the nonmass uptake lesions, PPV was 36.2% (17/47) for segmental, 20.1% (77/384) for focal, 30.8% (4/13) for diffuse, and 4.3% (5/116) for regional or multiple regional distribution. PPV was 5.3% (5/94) for one view, 15.2% (32/210) for two views, 14.6% (13/89) for three views, and 45.4% (113/249) for four views showing the lesion. PPV was 14.0% (43/307) for mild, 22.4% (51/228) for moderate, and 64.8% (70/108) for marked uptake intensity. CONCLUSION. The MBI lexicon lesion descriptors are associated with likelihood of malignancy. PPV was higher for masses, lesions seen on multiple MBI views, and lesions with marked uptake intensity. Among nonmass uptake lesions, PPV was highest for those with segmental distribution. CLINICAL IMPACT. Insight into the likelihood of malignancy associated with the MBI lexicon descriptors can inform radiologists' interpretations and guide potential future incorporation of the MBI lexicon into the ACR BI-RADS Atlas.

Understanding Reader Variability: A 25-Radiologist Study on Liver Metastasis Detection at CT.

Background Substantial interreader variability exists for common tasks in CT imaging, such as detection of hepatic metastases. This variability can undermine patient care by leading to misdiagnosis. Purpose To determine the impact of interreader variability associated with (a) reader experience, (b) image navigation patterns (eg, eye movements, workstation interactions), and (c) eye gaze time at missed liver metastases on contrast-enhanced abdominal CT images. Materials and Methods In a single-center prospective observational trial at an academic institution between December 2020 and February 2021, readers were recruited to examine 40 contrast-enhanced abdominal CT studies (eight normal, 32 containing 91 liver metastases). Readers circumscribed hepatic metastases and reported confidence. The workstation tracked image navigation and eye movements. Performance was quantified by using the area under the jackknife alternative free-response receiver operator characteristic (JAFROC-1) curve and per-metastasis sensitivity and was associated with reader experience and image navigation variables. Differences in area under JAFROC curve were assessed with the Kruskal-Wallis test followed by the Dunn test, and effects of image navigation were assessed by using the Wilcoxon signed-rank test. Results Twenty-five readers (median age, 38 years; IQR, 31-45 years; 19 men) were recruited and included nine subspecialized abdominal radiologists, five nonabdominal staff radiologists, and 11 senior residents or fellows. Reader experience explained differences in area under the JAFROC curve, with abdominal radiologists demonstrating greater area under the JAFROC curve (mean, 0.77; 95% CI: 0.75, 0.79) than trainees (mean, 0.71; 95% CI: 0.69, 0.73) (P = .02) or nonabdominal subspecialists (mean, 0.69; 95% CI: 0.60, 0.78) (P = .03). Sensitivity was similar within the reader experience groups (P = .96). Image navigation variables that were associated with higher sensitivity included longer interpretation time (P = .003) and greater use of coronal images (P < .001). The eye gaze time was at least 0.5 and 2.0 seconds for 71% (266 of 377) and 40% (149 of 377) of missed metastases, respectively. Conclusion Abdominal radiologists demonstrated better discrimination for the detection of liver metastases on abdominal contrast-enhanced CT images. Missed metastases frequently received at least a brief eye gaze. Higher sensitivity was associated with longer interpretation time and greater use of liver display windows and coronal images. © RSNA, 2022 Online supplemental material is available for this article.

Vessel wall imaging features of Moyamoya disease in a North American population: patterns of negative remodelling, contrast enhancement, wall thickening, and stenosis.

BACKGROUND: This study characterized vessel wall imaging (VWI) features of Moyamoya disease (MMD) in a predominantly adult population at a North American center. METHODS: Consecutive patients with VWI were included. Twelve arterial segments were analyzed for wall thickening, degree and pattern of contrast enhancement, and remodeling. RESULTS: Overall, 286 segments were evaluated in 24 patients (mean age = 36.0 years [range = 1-58]). Of 172 affected segments, 163 (95%) demonstrated negative remodeling. Complete vessel wall obliteration was most frequent in the proximal M1 (17/48, 35%). Affected segments enhanced in 72/172 (42%) (n = 15 for grade II; n = 54 for concentric and n = 18 for eccentric); 20 of 24 (83%) patients had at least one enhancing segment. Both enhancing and non-enhancing segments were present in 19/20 (95%) patients. Vessel wall enhancement was most common in the proximal segments and correlated to the degree of stenosis (p < 0.001), and outer wall diameter (p < 0.001), but not disease duration (p = 0.922) or Suzuki score (p = 0.477). Wall thickening was present in 82/172 (48%) affected segments and was associated with contrast enhancement (p < 0.001), degree of stenosis (p < 0.001), and smaller outer wall diameter (p = 0.004). CONCLUSION: This study presents VWI findings in North American patients with MMD. Negative remodeling was the most common finding. Most patients had both enhancing and non-enhancing abnormal segments. Vessel wall enhancement was most common in proximal segments, variable in pattern or degree and was correlated to the degree of stenosis and smaller outer wall diameter.

Zeta-Carotene Isomerase (Z-ISO) Is Required for Light-Independent Carotenoid Biosynthesis in the Cyanobacterium Synechocystis sp. PCC 6803.

Carotenoids are crucial photosynthetic pigments utilized for light harvesting, energy transfer, and photoprotection. Although most of the enzymes involved in carotenoid biosynthesis in chlorophototrophs are known, some are yet to be identified or fully characterized in certain organisms. A recently characterized enzyme in oxygenic phototrophs is 15-cis-zeta(ζ)-carotene isomerase (Z-ISO), which catalyzes the cis-to-trans isomerization of the central 15-15' cis double bond in 9,15,9'-tri-cis-ζ-carotene to produce 9,9'-di-cis-ζ-carotene during the four-step conversion of phytoene to lycopene. Z-ISO is a heme B-containing enzyme best studied in angiosperms. Homologs of Z-ISO are present in organisms that use the multi-enzyme poly-cis phytoene desaturation pathway, including algae and cyanobacteria, but appear to be absent in green bacteria. Here we confirm the identity of Z-ISO in the model unicellular cyanobacterium Synechocystis sp. PCC 6803 by showing that the protein encoded by the slr1599 open reading frame has ζ-carotene isomerase activity when produced in Escherichia coli. A Synechocystis Δslr1599 mutant synthesizes a normal quota of carotenoids when grown under illumination, where the photolabile 15-15' cis double bond of 9,15,9'-tri-cis-ζ-carotene is isomerized by light, but accumulates this intermediate and fails to produce 'mature' carotenoid species during light-activated heterotrophic growth, demonstrating the requirement of Z-ISO for carotenoid biosynthesis during periods of darkness. In the absence of a structure of Z-ISO, we analyze AlphaFold models of the Synechocystis, Zea mays (maize), and Arabidopsis thaliana enzymes, identifying putative protein ligands for the heme B cofactor and the substrate-binding site.