Hematopoietic Stem Cells Count and Remember Self-Renewal Divisions.

The ability of cells to count and remember their divisions could underlie many alterations that occur during development, aging, and disease. We tracked the cumulative divisional history of slow-cycling hematopoietic stem cells (HSCs) throughout adult life. This revealed a fraction of rarely dividing HSCs that contained all the long-term HSC (LT-HSC) activity within the aging HSC compartment. During adult life, this population asynchronously completes four traceable symmetric self-renewal divisions to expand its size before entering a state of dormancy. We show that the mechanism of expansion involves progressively lengthening periods between cell divisions, with long-term regenerative potential lost upon a fifth division. Our data also show that age-related phenotypic changes within the HSC compartment are divisional history dependent. These results suggest that HSCs accumulate discrete memory stages over their divisional history and provide evidence for the role of cellular memory in HSC aging.

How auditory neurons count temporal intervals and decode information.

The numerical sense of animals includes identifying the numerosity of a sequence of events that occur with specific intervals, e.g., notes in a call or bar of music. Across nervous systems, the temporal patterning of spikes can code these events, but how this information is decoded (counted) remains elusive. In the anuran auditory system, temporal information of this type is decoded in the midbrain, where "interval-counting" neurons spike only after at least a threshold number of sound pulses have occurred with specific timing. We show that this decoding process, i.e., interval counting, arises from integrating phasic, onset-type and offset inhibition with excitation that augments across successive intervals, possibly due to a progressive decrease in "shunting" effects of inhibition. Because these physiological properties are ubiquitous within and across central nervous systems, interval counting may be a general mechanism for decoding diverse information coded/encoded in temporal patterns of spikes, including "bursts," and estimating elapsed time.

Challenges and approaches to calibrating patient phenotype as evidence for cancer gene variant classification under ACMG/AMP guidelines.

Since first publication of the American College of Medical Genetics and Genomics/Association for Medical Pathology (ACMG/AMP) variant classification guidelines, additional recommendations for application of certain criteria have been released (https://clinicalgenome.org/docs/), to improve their application in the diagnostic setting. However, none have addressed use of the PS4 and PP4 criteria, capturing patient presentation as evidence towards pathogenicity. Application of PS4 can be done through traditional case-control studies, or "proband counting" within or across clinical testing cohorts. Review of the existing PS4 and PP4 specifications for Hereditary Cancer Gene Variant Curation Expert Panels revealed substantial differences in the approach to defining specifications. Using BRCA1, BRCA2 and TP53 as exemplar genes, we calibrated different methods proposed for applying the "PS4 proband counting" criterion. For each approach, we considered limitations, non-independence with other ACMG/AMP criteria, broader applicability, and variability in results for different datasets. Our findings highlight inherent overlap of proband-counting methods with ACMG/AMP frequency codes, and the importance of calibration to derive dataset-specific code weights that can account for potential between-dataset differences in ascertainment and other factors. Our work emphasizes the advantages and generalizability of logistic regression analysis over simple proband-counting approaches to empirically determine the relative predictive capacity and weight of various personal clinical features in the context of multigene panel testing, for improved variant interpretation. We also provide a general protocol, including instructions for data formatting and a web-server for analysis of personal history parameters, to facilitate dataset-specific calibration analyses required to use such data for germline variant classification.

TMBstable: a variant caller controls performance variation across heterogeneous sequencing samples.

In cancer genomics, variant calling has advanced, but traditional mean accuracy evaluations are inadequate for biomarkers like tumor mutation burden, which vary significantly across samples, affecting immunotherapy patient selection and threshold settings. In this study, we introduce TMBstable, an innovative method that dynamically selects optimal variant calling strategies for specific genomic regions using a meta-learning framework, distinguishing it from traditional callers with uniform sample-wide strategies. The process begins with segmenting the sample into windows and extracting meta-features for clustering, followed by using a pre-trained meta-model to select suitable algorithms for each cluster, thereby addressing strategy-sample mismatches, reducing performance fluctuations and ensuring consistent performance across various samples. We evaluated TMBstable using both simulated and real non-small cell lung cancer and nasopharyngeal carcinoma samples, comparing it with advanced callers. The assessment, focusing on stability measures, such as the variance and coefficient of variation in false positive rate, false negative rate, precision and recall, involved 300 simulated and 106 real tumor samples. Benchmark results showed TMBstable's superior stability with the lowest variance and coefficient of variation across performance metrics, highlighting its effectiveness in analyzing the counting-based biomarker. The TMBstable algorithm can be accessed at https://github.com/hello-json/TMBstable for academic usage only.

Digital microfluidics-based digital counting of single-cell copy number variation (dd-scCNV Seq).

Single-cell copy number variations (CNVs), major dynamic changes in humans, result in differential levels of gene expression and account for adaptive traits or underlying disease. Single-cell sequencing is needed to reveal these CNVs but has been hindered by single-cell whole-genome amplification (scWGA) bias, leading to inaccurate gene copy number counting. In addition, most of the current scWGA methods are labor intensive, time-consuming, and expensive with limited wide application. Here, we report a unique single-cell whole-genome library preparation approach based on digital microfluidics for digital counting of single-cell Copy Number Variation (dd-scCNV Seq). dd-scCNV Seq directly fragments the original single-cell DNA and uses these fragments as templates for amplification. These reduplicative fragments can be filtered computationally to generate the original partitioned unique identified fragments, thereby enabling digital counting of copy number variation. dd-scCNV Seq showed an increase in uniformity in the single-molecule data, leading to more accurate CNV patterns compared to other methods with low-depth sequencing. Benefiting from digital microfluidics, dd-scCNV Seq allows automated liquid handling, precise single-cell isolation, and high-efficiency and low-cost genome library preparation. dd-scCNV Seq will accelerate biological discovery by enabling accurate profiling of copy number variations at single-cell resolution.

When the Heart Meets the Mind: Exploring the Brain-Heart Interaction during Time Perception.

Recent studies suggest that time estimation relies on bodily rhythms and interoceptive signals. We provide the first direct electrophysiological evidence suggesting an association between the brain's processing of heartbeat and duration judgment. We examined heartbeat-evoked potential (HEP) and contingent negative variation (CNV) during an auditory duration-reproduction task and a control reaction-time task spanning 4, 8, and 12 s intervals, in both male and female participants. Interoceptive awareness was assessed with the Self-Awareness Questionnaire (SAQ) and interoceptive accuracy through the heartbeat-counting task (HCT). Results revealed that SAQ scores, but not the HCT, correlated with mean reproduced durations with higher SAQ scores associating with longer and more accurate duration reproductions. Notably, the HEP amplitude changes during the encoding phase of the timing task, particularly within 130-270 ms (HEP1) and 470-520 ms (HEP2) after the R-peak, demonstrated interval-specific modulations that did not emerge in the control task. A significant ramp-like increase in HEP2 amplitudes occurred during the duration-encoding phase of the timing but not during the control task. This increase within the reproduction phase of the timing task correlated significantly with the reproduced durations for the 8 s and the 4 s intervals. The larger the increase in HEP2, the greater the under-reproduction of the estimated duration. CNV components during the encoding phase of the timing task were more negative than those in the reaction-time task, suggesting greater executive resources orientation toward time. We conclude that interoceptive awareness (SAQ) and cortical responses to heartbeats (HEP) predict duration reproductions, emphasizing the embodied nature of time.

BIOMAPP::CHIP: large-scale motif analysis.

BACKGROUND: Discovery biological motifs plays a fundamental role in understanding regulatory mechanisms. Computationally, they can be efficiently represented as kmers, making the counting of these elements a critical aspect for ensuring not only the accuracy but also the efficiency of the analytical process. This is particularly useful in scenarios involving large data volumes, such as those generated by the ChIP-seq protocol. Against this backdrop, we introduce BIOMAPP::CHIP, a tool specifically designed to optimize the discovery of biological motifs in large data volumes. RESULTS: We conducted a comprehensive set of comparative tests with state-of-the-art algorithms. Our analyses revealed that BIOMAPP::CHIP outperforms existing approaches in various metrics, excelling both in terms of performance and accuracy. The tests demonstrated a higher detection rate of significant motifs and also greater agility in the execution of the algorithm. Furthermore, the SMT component played a vital role in the system's efficiency, proving to be both agile and accurate in kmer counting, which in turn improved the overall efficacy of our tool. CONCLUSION: BIOMAPP::CHIP represent real advancements in the discovery of biological motifs, particularly in large data volume scenarios, offering a relevant alternative for the analysis of ChIP-seq data and have the potential to boost future research in the field. This software can be found at the following address: (https://github.com/jadermcg/biomapp-chip).

Quantifying bunch-mode influence on photon-counting detectors at SPring-8.

Count-loss characteristics of photon-counting 2D detectors are demonstrated for eight bunch-modes at SPring-8 through Monte Carlo simulations. As an indicator, the effective maximum count rate was introduced to signify the X-ray intensity that the detector can count with a linearity of 1% or better after applying a count-loss correction in each bunch-mode. The effective maximum count rate is revealed to vary depending on the bunch-mode and the intrinsic dead time of the detectors, ranging from 0.012 to 0.916 Mcps (megacounts per second) for a 120 ns dead time, 0.009 to 0.807 Mcps for a 0.5 µs dead time and 0.020 to 0.273 Mcps for a 3 µs intrinsic detector dead time. Even with equal-interval bunch-modes at SPring-8, the effective maximum count rate does not exceed 1 Mcps pixel-1. In other words, to obtain data with a linearity better than 1%, the maximum intensity of X-rays entering the detector should be reduced to 1 Mcps pixel-1 or less, and, in some cases, even lower, depending on the bunch-mode. When applying count-loss correction using optimized dead times tailored to each bunch-mode, the effective maximum count rate exceeds the values above. However, differences in the effective maximum count rate due to bunch-modes persist. Users of photon-counting 2D detectors are encouraged to familiarize themselves with the count-loss characteristics dependent on bunch-mode, and to conduct experiments accordingly. In addition, when designing the time structure of bunch-modes at synchrotron radiation facilities, it is essential to take into account the impact on experiments using photon-counting 2D detectors.

The histomorphological and stereological assessment of rat dorsal root ganglion tissues after various types of sciatic nerve injury.

Peripheral nerve injuries lead to significant changes in the dorsal root ganglia, where the cell bodies of the damaged axons are located. The sensory neurons and the surrounding satellite cells rearrange the composition of the intracellular organelles to enhance their plasticity for adaptation to changing conditions and response to injury. Meanwhile, satellite cells acquire phagocytic properties and work with macrophages to eliminate degenerated neurons. These structural and functional changes are not identical in all injury types. Understanding the cellular response, which varies according to the type of injury involved, is essential in determining the optimal method of treatment. In this research, we investigated the numerical and morphological changes in primary sensory neurons and satellite cells in the dorsal root ganglion 30 days following chronic compression, crush, and transection injuries using stereology, high-resolution light microscopy, immunohistochemistry, and behavioral analysis techniques. Electron microscopic methods were employed to evaluate fine structural alterations in cells. Stereological evaluations revealed no statistically significant difference in terms of mean sensory neuron numbers (p > 0.05), although a significant decrease was observed in sensory neuron volumes in the transection and crush injury groups (p < 0.05). Active caspase-3 immunopositivity increased in the injury groups compared to the sham group (p < 0.05). While crush injury led to desensitization, chronic compression injury caused thermal hyperalgesia. Macrophage infiltrations were observed in all injury types. Electron microscopic results revealed that the chromatolysis response was triggered in the sensory neuron bodies from the transection injury group. An increase in organelle density was observed in the perikaryon of sensory neurons after crush-type injury. This indicates the presence of a more active regeneration process in crush-type injury than in other types. The effect of chronic compression injury is more devastating than that of crush-type injury, and the edema caused by compression significantly inhibits the regeneration process.

A longitudinal study of the role of fingers in the development of early number and arithmetic skills in children with Apert syndrome.

This paper discusses a longitudinal study with children with Apert syndrome aged between 4 and 11 years. There has long been an interest in the role of fingers in the development of early number skills and arithmetic. As children with Apert syndrome are born with complex fusions of their fingers, they have to undergo several surgical procedures in order to obtain individuated fingers. This has implications for their finger mobility and finger awareness. It has been suggested that children with Apert syndrome have specific difficulties with early number and arithmetic activities. The findings from this study suggest that engaging children with Apert syndrome in activities that develop finger awareness (finger gnosis) and finger mobility (fine motor skills) may have a positive impact on their ability to engage with appropriate mathematics curricula at school. This is relevant to all those involved in the care of children with Apert syndrome and will be of particular relevance to those involved in early childhood and primary education. This study also provides new insights into the role of finger use in the development of skills and understanding in early number and arithmetic.

Assessment and comparison of viability assays for cellular products.

BACKGROUND AIMS: Accurate assessment of cell viability is crucial in cellular product manufacturing, yet selecting the appropriate viability assay presents challenges due to various factors. This study compares and evaluates different viability assays on fresh and cryopreserved cellular products, including peripheral blood stem cell (PBSC) and peripheral blood mononuclear cell (PBMC) apheresis products, purified PBMCs and cultured chimeric antigen receptor and T-cell receptor-engineered T-cell products. METHODS: Viability assays, including manual Trypan Blue exclusion, flow cytometry-based assays using 7-aminoactinomycin D (7-AAD) or propidium iodide (PI) direct staining or cell surface marker staining in conjunction with 7-AAD, Cellometer (Nexcelom Bioscience LLC, Lawrence, MA, USA) Acridine Orange/PI staining and Vi-CELL BLU Cell Viability Analyzer (Beckman Coulter, Inc, Brea, CA, USA), were evaluated. A viability standard was established using live and dead cell mixtures to assess the accuracy of these assays. Furthermore, precision assessment was conducted to determine the reproducibility of the viability assays. Additionally, the viability of individual cell populations from cryopreserved PBSC and PBMC apheresis products was examined. RESULTS: All methods provided accurate viability measurements and generated consistent and reproducible viability data. The assessed viability assays were demonstrated to be reliable alternatives when evaluating the viability of fresh cellular products. However, cryopreserved products exhibited variability among the tested assays. Additionally, analyzing the viability of each subset of the cryopreserved PBSC and PBMC apheresis products revealed that T cells and granulocytes were more susceptible to the freeze-thaw process, showing decreased viability. CONCLUSIONS: The study demonstrates the importance of careful assay selection, validation and standardization, particularly for assessing the viability of cryopreserved products. Given the complexity of cellular products, choosing a fit-for-purpose viability assay is essential.

Optimizing retinal ganglion cell nuclear staining for automated cell counting.

The retinal ganglion cells (RGCs) serve as the critical pathway for transmitting visual information from the retina to the brain, yet they can be dramatically impacted by diseases such as glaucoma. When investigating disease processes affecting RGCs in mouse models, accurately quantifying affected cells becomes essential. However, the use of pan RGC markers like RBPMS or THY1 presents challenges in accurate total cell counting. While Brn3a serves as a reliable RGC nuclear marker for automated counting, it fails to encompass all RGC subtypes in mice. To address this limitation and enable precise automated counting, our research endeavors to develop a method for labeling nuclei in all RGC subtypes. Investigating RGC subtypes labeled with the nuclear marker POU6F2 revealed that numerous RGCs unlabeled by Brn3a were, in fact, labeled with POU6F2. We hypothesize that using antibodies against both Brn3a and POU6F2 would label virtually all RGC nuclei in the mouse retina. Our experiments confirmed that staining retinas with both markers resulted in the labeling of all RGCs. Additionally, when using the cell body marker RBPMS known to label all mouse RGCs, all RBPMS-labeled cells also exhibited Brn3a or POU6F2 labeling. This combination of Brn3a and POU6F2 antibodies provides a pan-RGC nuclear stain, facilitating accurate automated counting by labeling cell nuclei in the retina.

Cellaca® PLX image cytometer as an alternative for immunophenotyping, GFP/RFP transfection efficiencies, and apoptosis analysis.

Cell and gene therapy is a fast-growing field for cancer therapeutics requiring reliable instrumentation and technologies. Key parameters essential for satisfying Chemistry Manufacturing and Controls criteria standards are routinely performed using flow cytometry. Recently, image cytometry was developed for cell characterization and cell-based assays but had not yet demonstrated sufficient sensitivity for surface marker detection. We developed the Cellaca® PLX image cytometry system and the respective methodologies required for immunophenotyping, GFP and RFP transfection/transduction efficiencies, and cell health analyses for routine cell characterization. All samples tested were compared directly to results from the CytoFLEX flow cytometer. PBMCs were stained with T-cell surface markers for immunophenotyping, and results show highly comparable CD3, CD4, and CD8 populations (within 5 %). GFP- or RFP-expressing cell lines were analyzed for transfection/transduction efficiencies, and the percentage positive cells and respective viabilities were equivalent on both systems. Staurosporine-treated Jurkat cells were stained for apoptotic markers, where annexin V and caspase-3 positive cells were within 5 % comparing both instruments. The proposed system may provide a complementary tool for performing routine cell-based experiments with improved efficiency and sensitivity compared to prior image cytometers, which may be significantly valuable to the cell and gene therapy field.

Looks like home: numerosity, but not spatial frequency guides preference in zebrafish larvae (Danio rerio).

Despite their young age, zebrafish larvae have a well-developed visual system and can distinguish between different visual stimuli. First, we investigated if the first visual surroundings the larvae experience during the first days after hatching shape their habitat preference. Indeed, these animals seem to "imprint" on the first surroundings they see and select visual stimuli accordingly at 7 days post fertilization (dpf). In particular, if zebrafish larvae experience a bar background just after hatching, they later on prefer bars over white stimuli, and vice versa. We then used this acquired preference for bars to investigate innate numerical abilities. We wanted to specifically test if the zebrafish larvae show real numerical abilities or if they rely on a lower-level mechanism-i.e. spatial frequency-to discriminate between two different numerosities. When we matched the spatial frequency in stimuli with different numbers of bars, the larvae reliably selected the higher numerosity. A previous study has ruled out that 7 dpf zebrafish larvae use convex hull, cumulative surface area and density to choose between two numerosities. Therefore, our results indicate that zebrafish larvae rely on real numerical abilities rather than other cues, including spatial frequency, when spontaneously comparing two sets with different numbers of bars.

Comparison of automated kidney stone size measurement and volumetry in photon counting CT compared to 3rd generation dual energy CT and physically measurements - an ex vivo study.

PURPOSE: This ex vivo study aimed to compare a newly developed dual-source photon-counting CT (PCCT) with a 3rd generation dual-source dual-energy CT (DECT) for the detection and measurement (stone lengths and volumetrics) of urinary stones. METHODS: 143 urinary stones with a known geometry were physically measured and defined as reference values. Next, urinary stones were placed in an anthropomorphic abdomen-model and were scanned with DECT and PCCT. Images were read by two experienced examiners and automatically evaluated using a specific software. RESULTS: DECT and PCCT showed a high sensitivity for manual stone detection of 97.9% and 94.4%, and for automatic detection of 93.0% and 87.4%, respectively. Compared to that uric acid and xanthine stones were recognized slightly worse by DECT and PCCT with manual stone detection (93.3% and 82.2%), and with automatic detection (77.8% and 60.0%). All other stone entities were completely recognized. By comparing the maximum diameter of the reference value and DECT, Pearson-correlation was 0.96 (p < 0.001) for manual and 0.97 (p < 0.001) for automatic measurement, and for PCCT it was 0.94 (p < 0.001) for manual and 0.97 (p < 0.001) for automatic measurements. DECT and PCCT can also reliably determine volume manually and automatically with a Pearson-correlation of 0.99 (p < 0.001), respectively. CONCLUSION: Both CTs showed comparable results in stone detection, length measurement and volumetry compared to the reference values. Automatic measurement tends to underestimate the maximum diameter. DECT proved to be slightly superior in the recognition of xanthine and uric acid stones.

Image quality of lung perfusion with photon-counting-detector CT: comparison with dual-source, dual-energy CT.

PURPOSE: To evaluate the quality of lung perfusion imaging obtained with photon-counting-detector CT (PCD-CT) in comparison with dual-source, dual-energy CT (DECT). METHODS: Seventy-one consecutive patients scanned with PCD-CT were compared to a paired population scanned with dual-energy on a 3rd-generation DS-CT scanner using (a) for DS-CT (Group 1): collimation: 64 × 0.6 × 2 mm; pitch: 0.55; (b) for PCD-CT (Group 2): collimation: 144 × 0.4 mm; pitch: 1.5; single-source acquisition. The injection protocol was similar in both groups with the reconstruction of perfusion images by subtraction of high- and low-energy virtual monoenergetic images. RESULTS: Compared to Group 1, Group 2 examinations showed: (a) a shorter duration of data acquisition (0.93 ± 0.1 s vs 3.98 ± 0.35 s; p < 0.0001); (b) a significantly lower dose-length-product (172.6 ± 55.14 vs 339.4 ± 75.64 mGy·cm; p < 0.0001); and (c) a higher level of objective noise (p < 0.0001) on mediastinal images. On perfusion images: (a) the mean level of attenuation did not differ (p = 0.05) with less subjective image noise in Group 2 (p = 0.049); (b) the distribution of scores of fissure visualization differed between the 2 groups (p < 0.0001) with a higher proportion of fissures sharply delineated in Group 2 (n = 60; 84.5% vs n = 26; 26.6%); (c) the rating of cardiac motion artifacts differed between the 2 groups (p < 0.0001) with a predominance of examinations rated with mild artifacts in Group 2 (n = 69; 97.2%) while the most Group 1 examinations showed moderate artifacts (n = 52; 73.2%). CONCLUSION: PCD-CT acquisitions provided similar morphologic image quality and superior perfusion imaging at lower radiation doses. CLINICAL RELEVANCE STATEMENT: The improvement in the overall quality of perfusion images at lower radiation doses opens the door for wider applications of lung perfusion imaging in clinical practice. KEY POINTS: The speed of data acquisition with PCD-CT accounts for mild motion artifacts. Sharply delineated fissures are depicted on PCD-CT perfusion images. High-quality perfusion imaging was obtained with a 52% dose reduction.

Synthetic hematocrit from virtual non-contrast images for myocardial extracellular volume evaluation with photon-counting detector CT.

OBJECTIVES: To assess the accuracy of a synthetic hematocrit derived from virtual non-contrast (VNC) and virtual non-iodine images (VNI) for myocardial extracellular volume (ECV) computation with photon-counting detector computed tomography (PCD-CT). MATERIALS AND METHODS: Consecutive patients undergoing PCD-CT including a coronary CT angiography (CCTA) and a late enhancement (LE) scan and having a blood hematocrit were retrospectively included. In the first 75 patients (derivation cohort), CCTA and LE scans were reconstructed as VNI at 60, 70, and 80 keV and as VNC with quantum iterative reconstruction (QIR) strengths 2, 3, and 4. Blood pool attenuation (BPmean) was correlated to blood hematocrit. In the next 50 patients (validation cohort), synthetic hematocrit was calculated using BPmean. Myocardial ECV was computed using the synthetic hematocrit and compared with the ECV using the blood hematocrit as a reference. RESULTS: In the derivation cohort (49 men, mean age 79 ± 8 years), a correlation between BPmean and blood hematocrit ranged from poor for VNI of CCTA at 80 keV, QIR2 (R2 = 0.12) to moderate for VNI of LE at 60 keV, QIR4; 70 keV, QIR3 and 4; and VNC of LE, QIR3 and 4 (all, R2 = 0.58). In the validation cohort (29 men, age 75 ± 14 years), synthetic hematocrit was calculated from VNC of the LE scan, QIR3. Median ECV was 26.9% (interquartile range (IQR), 25.5%, 28.8%) using the blood hematocrit and 26.8% (IQR, 25.4%, 29.7%) using synthetic hematocrit (VNC, QIR3; mean difference, -0.2%; limits of agreement, -2.4%, 2.0%; p = 0.33). CONCLUSION: Synthetic hematocrit calculated from VNC images enables an accurate computation of myocardial ECV with PCD-CT. CLINICAL RELEVANCE STATEMENT: Virtual non-contrast images from cardiac late enhancement scans with photon-counting detector CT allow the calculation of a synthetic hematocrit, which enables accurate computation of myocardial extracellular volume. KEY POINTS: Blood hematocrit is mandatory for conventional myocardial extracellular volume computation. Synthetic hematocrit can be calculated from virtual non-iodine and non-contrast photon-counting detector CT images. Synthetic hematocrit from virtual non-contrast images enables computation of the myocardial extracellular volume.

Diagnostic performance of photon-counting detector CT for differentiation between adrenal adenomas and metastases.

OBJECTIVES: Aim of this study was to assess the value of virtual non-contrast (VNC) reconstructions in differentiating between adrenal adenomas and metastases on a photon-counting detector CT (PCD-CT). MATERIAL AND METHODS: Patients with adrenal masses and contrast-enhanced CT scans in portal venous phase were included. Image reconstructions were performed, including conventional VNC (VNCConv) and PureCalcium VNC (VNCPC), as well as virtual monochromatic images (VMI, 40-90 keV) and iodine maps. We analyzed images using semi-automatic segmentation of adrenal lesions and extracted quantitative data. Logistic regression models, non-parametric tests, Bland-Altman plots, and a random forest classifier were used for statistical analyses. RESULTS: The final study cohort consisted of 90 patients (36 female, mean age 67.8 years [range 39-87]) with adrenal lesions (45 adenomas, 45 metastases). Compared to metastases, adrenal adenomas showed significantly lower CT-values in VNCConv and VNCPC (p = 0.007). Mean difference between VNC and true non-contrast (TNC) was 17.67 for VNCConv and 14.85 for VNCPC. Random forest classifier and logistic regression models both identified VNCConv and VNCPC as the best discriminators. When using 26 HU as the threshold in VNCConv reconstructions, adenomas could be discriminated from metastases with a sensitivity of 86.7% and a specificity of 75.6%. CONCLUSION: VNC algorithms overestimate CT values compared to TNC in the assessment of adrenal lesions. However, they allow a reliable discrimination between adrenal adenomas and metastases and could be used in clinical routine in near future with an increased threshold (e.g., 26 HU). Further (multi-center) studies with larger patient cohorts and standardized protocols are required. CLINICAL RELEVANCE STATEMENT: VNC reconstructions overestimate CT values compared to TNC. Using a different threshold (e.g., 26 HU compared to the established 10 HU), VNC has a high diagnostic accuracy for the discrimination between adrenal adenomas and metastases. KEY POINTS: • Virtual non-contrast reconstructions may be promising tools to differentiate adrenal lesions and might save further diagnostic tests. • The conventional and a new calcium-preserving virtual non-contrast algorithm tend to systematically overestimate CT-values compared to true non-contrast images. • Therefore, increasing the established threshold for true non-contrast images (e.g., 10HU) may help to differentiate between adrenal adenomas and metastases on contrast-enhanced CT.

Diagnosis of acute pulmonary embolism: when photon-counting-detector CT replaces energy-integrating-detector CT in daily routine.

PURPOSE: To compare the diagnostic approach of acute pulmonary embolism (PE) with photon-counting-detector CT (PCD-CT) and energy-integrating-detector CT (EID-CT). MATERIALS AND METHODS: Two cohorts underwent CT angiographic examinations with EID-CT (Group 1; n = 158) and PCD-CT (Group 2; n = 172), (b) with two options in Group 1, dual energy (Group 1a) or single energy (Group 1b) and a single option in Group 2 (spectral imaging with single source). RESULTS: In Group 2, all patients benefited from spectral imaging, only accessible to 105 patients (66.5%) in Group 1, with a mean acquisition time significantly shorter (0.9 ± 0.1 s vs 4.0 ± 0 .3 s; p < 0.001) and mean values of CTDIvol and DLP reduced by 46.3% and 47.7%, respectively. Comparing the quality of 70 keV (Group 2) and averaged (Group 1a) images: (a) the mean attenuation within pulmonary arteries did not differ (p = 0.13); (b) the image noise was significantly higher (p < 0.001) in Group 2 with no difference in subjective image noise (p = 0.29); and (c) 89% of examinations were devoid of artifacts in Group 2 vs 28.6% in Group 1a. The percentage of diagnostic examinations was 95.2% (100/105; Group 1a), 100% (53/53; Group 1b), and 95.3% (164/172; Group 2). There were 4.8% (5/105; Group 1a) and 4.7% (8/172; Group 2) of non-diagnostic examinations, mainly due to the suboptimal quality of vascular opacification with the restoration of a diagnostic image quality on low-energy images. CONCLUSION: Compared to EID-CT, morphology and perfusion imaging were available in all patients scanned with PCD-CT, with the radiation dose reduced by 48%. CLINICAL RELEVANCE STATEMENT: PCD-CT enables scanning patients with the advantages of both spectral imaging, including high-quality morphologic imaging and lung perfusion for all patients, and fast scanning-a combination that is not simultaneously accessible with EID-CT while reducing the radiation dose by almost 50%.

Evaluation of ECG-Gated, High-Pitch Thoracoabdominal Angiographies With Dual-Source Photon-Counting Detector Computed Tomography.

PURPOSE: The aim of this study was to evaluate the radiation dose, image quality, and the potential of virtual monoenergetic imaging (VMI) reconstructions of high-pitch computed tomography angiography (CTA) of the thoracoabdominal aorta on a dual-source photon-counting detector-CT (PCD-CT) in comparison with an energy-integrating detector-CT (EID-CT), with a special focus on low-contrast attenuation. METHODS: Consecutive patients being referred for an electrocardiogram (ECG)-gated, high-pitch CTA of the thoracoabdominal aorta prior to transcatheter aortic valve replacement (TAVR), and examined on the PCD-CT, were included in this prospective single-center study. For comparison, a retrospective patient group with ECG-gated, high-pitch CTA examinations of the thoracoabdominal aorta on EID-CT with a comparable scan protocol was matched for gender, body mass index, height, and age. Virtual monoenergetic imaging reconstructions from 40 to 120 keV were performed. Enhancement and noise were measured in 7 vascular segments and the surrounding air as mean and standard deviation of CT values. The radiation dose was noted and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. Finally, a subgroup analysis was performed, comparing VMI reconstructions from 40 keV to 70 keV in patients with at least a 50% decrease in contrast attenuation between the ascending aorta and femoral arteries. RESULTS: Fifty patients (mean age 77.0±14.5 years; 31 women) were included. The radiation dose was significantly lower on the PCD-CT (4.2±1.4 vs. 7.2±2.2 mGy; p<0.001). With increasing keV, vascular noise, SNR, and CNR decreased. Intravascular attenuation was significantly higher on VMI at levels from 40 to 65, compared with levels of 120 keV (p<0.01 and p<0.005, respectively). On the PCD-CT, SNR was significantly higher in keV levels 40 and 70 (all p<0.001), and CNR was higher at keV levels 40 and 45 (each p<0.001), compared with scans on the EID-CT. At VMI ≤60 keV, image noise was also significantly higher than that in the control group. The subgroup analysis showed a drastically improved diagnostic performance of the low-keV images in patients with low-contrast attenuation. CONCLUSION: The ECG-gated CTA of the thoracoabdominal aorta in high-pitch mode on PCD-CT have significantly lower radiation dose and higher objective image quality than EID-CT. In addition, low-keV VMI can salvage suboptimal contrast studies, further reducing radiation dose by eliminating the need for repeat scans. CLINICAL IMPACT: ECG-gated CT-angiographies of the thoracoabdominal aorta can be acquired with a lower radtiation dose and a better image quality by using a dual-source photon-countinge detector CT. Furthermore, the inherent spectral data offers the possiblity to improve undiagnostic images and thus saves the patient from further radiation and contrast application.