Engineering regioselectivity of glycosyltransferase for efficient polydatin synthesis.

Resveratrol is a promising functional ingredient applied in food products. However, low bioavailability and poor water solubility, which can be improved by glycosylation, hinder its application. A uridine diphosphate-dependent glycosyltransferase (UGT) from Bacillus subtilis 168 (named UGTBS) presents potential application for resveratrol glycosylation; nonetheless, imprecise regioselectivity renders the synthesis of resveratrol-3-O-ß-D-glucoside (polydatin) difficult. Therefore, molecular evolution was applied to UGTBS. A triple mutant Y14I/I62G/M315W was developed for 3-OH glycosylation of resveratrol and polydatin accounted for 91% of the total product. Kinetic determination and molecular docking indicated that the enhancement of hydrogen bond interaction and altered conformation of the binding pocket increases the enzyme's affinity for the 3-OH group, stabilizing the enzyme-substrate intermediate and promoting polydatin formation. Furthermore, a fed-batch cascade reaction by periodic addition of resveratrol was conducted and nearly 20 mM polydatin was obtained. The mutant Y14I/I62G/M315W can be used for polydatin manufacture.

Improvement in organic solvent resistance of keratinase BLk by directed evolution.

Keratinase, a vital enzyme in hair degradation, requires enhanced stability for industrial applications in the harsh reaction environment used for keratin hydrolysis. Previous studies have focused on improving keratinase thermostability. In this study, directed evolution was applied to enhance the organic solvent stability of the keratinase BLk from Bacillus licheniformis. Three mutants were identified, exhibiting significant enhanced stability in various solvents, although no similar improvements were observed in terms of thermostability. The identified mutations were located on the enzyme surface. The half-lives of the D41A, A24E, and A24Q mutants increased by 47-, 63-, and 61-fold, respectively, in the presence of 50% (v/v) acetonitrile compared to that of the wild type (WT). Similarly, in the presence of 50% (v/v) acetone, the half-lives of these mutants increased by 22-, 27-, and 27-fold compared to that of the WT enzyme. Notably, the proteolytic activity of all the selected mutants was similar to that of the WT enzyme. Furthermore, molecular dynamics simulation was used to assess the possible reasons for enhanced solvent stability. These results suggest that heightened intramolecular interactions, such as hydrogen bonding and hydrophobic interactions, contribute to improved solvent tolerance. The mutants obtained in this study hold significant potential for industrial applications.

Cardiac involvement in COVID-19 patients: mid-term follow up by cardiovascular magnetic resonance.

BACKGROUND: Coronavirus disease 2019 (COVID-19) induces myocardial injury, either direct myocarditis or indirect injury due to systemic inflammatory response. Myocardial involvement has been proved to be one of the primary manifestations of COVID-19 infection, according to laboratory test, autopsy, and cardiovascular magnetic resonance (CMR). However, the middle-term outcome of cardiac involvement after the patients were discharged from the hospital is yet unknown. The present study aimed to evaluate mid-term cardiac sequelae in recovered COVID-19 patients by CMR METHODS: A total of 47 recovered COVID-19 patients were prospectively recruited and underwent CMR examination. The CMR protocol consisted of black blood fat-suppressed T2 weighted imaging, T2 star mapping, left ventricle (LV) cine imaging, pre- and post-contrast T1 mapping, and late gadolinium enhancement (LGE). LGE were assessed in mixed both recovered COVID-19 patients and healthy controls. The LV and right ventricle (RV) function and LV mass were assessed and compared with healthy controls. RESULTS: A total of 44 recovered COVID-19 patients and 31 healthy controls were studied. LGE was found in 13 (30%) of COVID-19 patients. All LGE lesions were located in the mid myocardium and/or sub-epicardium with a scattered distribution. Further analysis showed that LGE-positive patients had significantly decreased LV peak global circumferential strain (GCS), RV peak GCS, RV peak global longitudinal strain (GLS) as compared to non-LGE patients (p < 0.05), while no difference was found between the non-LGE patients and healthy controls. CONCLUSION: Myocardium injury existed in 30% of COVID-19 patients. These patients have depressed LV GCS and peak RV strains at the 3-month follow-up. CMR can monitor the COVID-19-induced myocarditis progression, and CMR strain analysis is a sensitive tool to evaluate the recovery of LV and RV dysfunction.

Liver Venous Tree Separation via Twin-Line RANSAC and Murray's Law.

It is essential for physicians to obtain the accurate venous tree from abdominal CT angiography (CTA) series in order to carry out the preoperative planning and intraoperative navigation for hepatic surgery. In this process, one of the important tasks is to separate the given liver venous mask into its hepatic and portal parts. In this paper, we present a novel method for liver venous tree separation. The proposed method first concentrates on extracting potential vessel intersection points between hepatic and portal venous systems. Then, the proposed method focuses on modeling the vessel intersection neigh-borhoods with a robust twin-line random sample consensus (RANSAC) shape detector. Finally, the proposed method conducts the venous tree separation based on the results of the twin-line RANSAC as well as physical constraints posed by Murray's Law. We test our method on 22 clinical CTA series and demonstrate its effectiveness.

Inferior vena cava segmentation with parameter propagation and graph cut.

PURPOSE: The inferior vena cava (IVC) is one of the vital veins inside the human body. Accurate segmentation of the IVC from contrast-enhanced CT images is of great importance. This extraction not only helps the physician understand its quantitative features such as blood flow and volume, but also it is helpful during the hepatic preoperative planning. However, manual delineation of the IVC is time-consuming and poorly reproducible. METHODS: In this paper, we propose a novel method to segment the IVC with minimal user interaction. The proposed method performs the segmentation block by block between user-specified beginning and end masks. At each stage, the proposed method builds the segmentation model based on information from image regional appearances, image boundaries, and a prior shape. The intensity range and the prior shape for this segmentation model are estimated based on the segmentation result from the last block, or from user- specified beginning mask if at first stage. Then, the proposed method minimizes the energy function and generates the segmentation result for current block using graph cut. Finally, a backward tracking step from the end of the IVC is performed if necessary. RESULTS: We have tested our method on 20 clinical datasets and compared our method to three other vessel extraction approaches. The evaluation was performed using three quantitative metrics: the Dice coefficient (Dice), the mean symmetric distance (MSD), and the Hausdorff distance (MaxD). The proposed method has achieved a Dice of [Formula: see text], an MSD of [Formula: see text] mm, and a MaxD of [Formula: see text] mm, respectively, in our experiments. CONCLUSION: The proposed approach can achieve a sound performance with a relatively low computational cost and a minimal user interaction. The proposed algorithm has high potential to be applied for the clinical applications in the future.

Clinical evaluation of new automatic coronary-specific best cardiac phase selection algorithm for single-beat coronary CT angiography.

The aim of this study was to evaluate the workflow efficiency of a new automatic coronary-specific reconstruction technique (Smart Phase, GE Healthcare-SP) for selection of the best cardiac phase with least coronary motion when compared with expert manual selection (MS) of best phase in patients with high heart rate. A total of 46 patients with heart rates above 75 bpm who underwent single beat coronary computed tomography angiography (CCTA) were enrolled in this study. CCTA of all subjects were performed on a 256-detector row CT scanner (Revolution CT, GE Healthcare, Waukesha, Wisconsin, US). With the SP technique, the acquired phase range was automatically searched in 2% phase intervals during the reconstruction process to determine the optimal phase for coronary assessment, while for routine expert MS, reconstructions were performed at 5% intervals and a best phase was manually determined. The reconstruction and review times were recorded to measure the workflow efficiency for each method. Two reviewers subjectively assessed image quality for each coronary artery in the MS and SP reconstruction volumes using a 4-point grading scale. The average HR of the enrolled patients was 91.1±19.0bpm. A total of 204 vessels were assessed. The subjective image quality using SP was comparable to that of the MS, 1.45±0.85 vs 1.43±0.81 respectively (p = 0.88). The average time was 246 seconds for the manual best phase selection, and 98 seconds for the SP selection, resulting in average time saving of 148 seconds (60%) with use of the SP algorithm. The coronary specific automatic cardiac best phase selection technique (Smart Phase) improves clinical workflow in high heart rate patients and provides image quality comparable with manual cardiac best phase selection. Reconstruction of single-beat CCTA exams with SP can benefit the users with less experienced in CCTA image interpretation.

High-pitch, low-voltage and low-iodine-concentration CT angiography of aorta: assessment of image quality and radiation dose with iterative reconstruction.

OBJECTIVE: To assess the image quality of aorta obtained by dual-source computed tomography angiography (DSCTA), performed with high pitch, low tube voltage, and low iodine concentration contrast medium (CM) with images reconstructed using iterative reconstruction (IR). METHODS: One hundred patients randomly allocated to receive one of two types of CM underwent DSCTA with the electrocardiogram-triggered Flash protocol. In the low-iodine group, 50 patients received CM containing 270 mg I/mL and were scanned at low tube voltage (100 kVp). In the high-iodine CM group, 50 patients received CM containing 370 mg I/mL and were scanned at the tube voltage (120 kVp). The filtered back projection (FBP) algorithm was used for reconstruction in both groups. In addition, the IR algorithm was used in the low-iodine group. Image quality of the aorta was analyzed subjectively by a 3-point grading scale and objectively by measuring the CT attenuation in terms of the signal- and contrast-to-noise ratios (SNR and CNR, respectively). Radiation and CM doses were compared. RESULTS: The CT attenuation, subjective image quality assessment, SNR, and CNR of various aortic regions of interest did not differ significantly between two groups. In the low-iodine group, images reconstructed by FBP and IR demonstrated significant differences in image noise, SNR, and CNR (p<0.05). The low-iodine group resulted in 34.3% less radiation (4.4 ± 0.5 mSv) than the high-iodine group (6.7 ± 0.6 mSv), and 27.3% less iodine weight (20.36 ± 2.65 g) than the high-iodine group (28 ± 1.98 g). Observers exhibited excellent agreement on the aortic image quality scores (κ = 0.904). CONCLUSIONS: CT images of aorta could be obtained within 2 s by using a DSCT Flash protocol with low tube voltage, IR, and low-iodine-concentration CM. Appropriate contrast enhancement was achieved while maintaining good image quality and decreasing the radiation and iodine doses.

Assessment of right ventricular function with 320-slice volume cardiac CT: comparison with cardiac magnetic resonance imaging.

To evaluate the accuracy and feasibility of right ventricular function parameters measurement using 320-slice volume cardiac CT. Retrospective analysis of 50 consecutive patients (23 men, 27 women) with suspected pulmonary diseases was performed in electrocardiogram (ECG)-gated cardiac CT and cardiac magnetic resonance (CMR). Parameters including right ventricular end-diastolic volume (RVEDV), right ventricular end- systolic volume (RVESV), right ventricular stroke volume (RVSV), right ventricular cardiac output (RVCO), and right ventricular ejection fraction (RVEF) were semi-automatically and separately calculated from both CT and CMR data. Significant difference between measurements was measured by paired t test and two-variable linear regression analysis with Pearson's correlation coefficient. Bland-Altman analysis was performed in each pair of parameters. There was little variability between the measurements by the two observers (kappa = 0.895-0.980, P < 0.05). There was good correlation between all parameters obtained by CT and CMR (P < 0.001): RVEDV (108.5 ± 21.9 ml, 113.5 ± 24.8 ml, r = 0.944), RVESV (69.8 ± 33.4 ml, 73.2 ± 35.4 ml, r = 0.972), RVSV (39.0 ± 13.2 ml, 40.2 ± 13.3 ml, r = 0.977), RVCO (2.6 ± 0.7 l, 2.6 ± 0.7 l. r = 0.958), RVEF (38.8 ± 19.1 %, 39.1 ± 19.3 %, r = 0.990), and there was no significant difference between CT and CMR measurements in RVEF (n = 50, t = -0.677, P > 0.05). 320-slice volume cardiac CT is an accurate non-invasive technique to evaluate RV function.

Low dose 320-row CT for left atrium and pulmonary veins imaging--the feasibility study.

OBJECTIVE: To evaluate the feasibility of low dose target-CTA volume scan for left atrium and pulmonary veins imaging using 320-row CT. METHODS: Forty-two patients (females 12, males 30; mean age 55.2 years; mean body mass index (BMI) 25.7 kg/m(2)) with persistent or intermittent atrial fibrillation before catheter ablation were enrolled in this study. Scan protocol was target-CTA volume scan under prospective ECG-gating. The target of the exposure was only set at 40% of the R-R interval (which was at the left atrium maximal volume). The exposure time was minimal setting (350 ms). Tube voltage 100 kV (BMI ≤ 25 kg/m(2)) or 120 kV (BMI>25 kg/m(2)), tube current 350-420 mA, and field of view of 180 mm × 180 mm-200 mm × 200 mm were used. The scanning range was from the level of the tracheal bifurcation to the diaphragm, and enabled to cover the left atrium and central pulmonary veins (120-140 mm). All of patients were divided into two groups according kV setting (100 kV and 120 kV). The image quality (good, moderate and poor) was evaluated by two reviewers. The CT-attenuation, image noise and contrast to noise ratio (CNR) of left atrium and every pulmonary vein branch were evaluated, respectively. The effective dose (ED) was calculated using a conversion coefficient for the chest 0.017. RESULTS: Of 42 patients, the image quality was good in 26 cases (62%) and moderate in 16 cases (38%). There was no statistical difference in the CT-attenuation and CNR of left atrium and central pulmonary veins between the two groups (P>0.05). The mean ED was 1.90 ± 0.19 mSv (range 1.57-2.25 mSv) in 100 kV group, and 3.83 ± 0.31 mSv (range 3.39-4.54 mSv) in 120 kV group. CONCLUSIONS: The low dose target-CTA volume scan is feasible in pulmonary veins and left atrium imaging using 320-row CT. Slim patients can be scanned at 100 kV setting without loss of image quality.

320-row CT coronary angiography: effect of 100-kV tube voltages on image quality, contrast volume, and radiation dose.

To prospectively evaluate image quality parameters, contrast volume and radiation dose at the 100-kilovolt (kV) setting during coronary computed tomographic angiography (CCTA) on a 320-row computed tomography scanner. We enrolled 107 consecutive patients with a heart rate <65 beats per minute (bpm) undergoing prospective electrocardiogram (ECG)-triggered CCTA. Forty patients with a body mass index (BMI) <25 kg/m(2) were scanned using 100-kV tube voltage settings, while 67 patients were scanned using 120-kV protocols. Image quality was assessed by two readers unaware of patient information and scan parameters. Attenuation in the aorta and perivascular fat tissue and image noise were measured. Contrast-to-noise ratios (CNRs) and contrast material volumes were calculated. The effective radiation doses were estimated using a chest conversion coefficient (0.017). Diagnostic image quality was achieved in 98.2% of coronary segments with 100-kV CCTA and 98.6% of coronary segments with 120-kV CCTA, with no significant differences in image quality scores for each coronary segment. Vessel attenuation, image noise, and CNR were not significantly different between the 100- and 120-kV protocols. Mean contrast injection rate and mean material volume were significantly lower for the 100-kV CCTA (4.35 ± 0.28 ml/s and 53.13 ± 3.77 ml, respectively) than for the 120-kV CCTA (5.16 ± 0.21 ml/s and 62.40 ± 3.66 ml respectively; P < 0.001). The effective radiation dose was 2.12 ± 0.19 mSv for 100-kV CCTA, a reduction of 54% compared to 4.61 ± 0.82 mSv for 120-kV CCTA. A 100-kV CCTA can be implemented in patients with a BMI < 25 kg/m(2). The 100-kV setting allows significant reductions in contrast material volume and effective radiation dose while maintaining adequate diagnostic image quality.

64-MDCT coronary angiography of patients with atrial fibrillation: influence of heart rate on image quality and efficacy in evaluation of coronary artery disease.

OBJECTIVE: The purpose of this study was to prospectively evaluate the effects of mean heart rate and heart rate variability on image quality at coronary 64-MDCT angiography of patients with atrial fibrillation and to determine the efficacy of coronary 64-MDCT angiography in the detection of significant (> 50%) coronary artery stenosis in patients with atrial fibrillation. SUBJECTS AND METHODS: Sixty patients (37 women, 23 men; mean age, 58.7 years) underwent both coronary 64-MDCT angiography and conventional coronary angiography. Heart rate variability was calculated as the SD from the mean heart rate during scanning. Image quality (good, moderate, or poor) and the presence of significant (> 50%) stenosis at coronary CT angiography were evaluated by two radiologists blinded to the results of conventional coronary angiography. The sensitivity, specificity, positive predictive value, and negative predictive value of coronary CT angiography were calculated with conventional angiography as the reference standard. Pearson's correlation analysis and chi-square tests were performed to compare image quality with mean heart rate and heart rate variability in each patient. Differences in detection of coronary artery stenosis between coronary 64-MDCT angiography and conventional coronary angiography were evaluated with McNemar's test, and agreement between techniques was calculated with kappa statistics. RESULTS: The mean heart rate was 90 +/- 13.1 beats/min with variability of 19.35 +/- 6.95 beats/min. Of 803 segments evaluated, 26 (3%) were considered to have poor image quality. Highly significant correlation was found between mean heart rate and image quality for all segments in each patient (r = 0.51, p < 0.001), for the right coronary artery (r = 0.43, p = 0.001), and for the distal portion of the coronary arteries (r = 0.50, p < 0.001). Heart rate variability also correlated in a highly significant way with image quality in all segments (r = 0.57, p < 0.001), the right coronary artery (r = 0.46, p < 0.001), and the middle (r = 0.44, p = 0.001) and distal (r = 0.41, p = 0.001) portions of the coronary arteries. The best diagnostic image quality was obtained in end systole (200-400 milliseconds). Image quality decreased significantly with a mean heart rate greater than 100 beats/min or with an SD of heart rate greater than 24 beats/min. The overall sensitivity, specificity, positive predictive value, and negative predictive value per segment level were 86.4%, 99.3%, 79.2%, and 99.6%. No significant difference was found between coronary 64-MDCT angiography and conventional coronary angiography in detection of significant stenosis. Excellent agreement between techniques was found. CONCLUSION: Coronary 64-MDCT angiography has diagnostic image quality within a wide range of heart rates and in patients with atrial fibrillation. Reducing average heart rate and heart rate variability in patients with atrial fibrillation is beneficial in improving image quality.

Hemorrhage and large lipid-rich necrotic cores are independently associated with thin or ruptured fibrous caps: an in vivo 3T MRI study.

OBJECTIVE: Histological studies suggest associations between hemorrhage and large lipid-rich/necrotic cores with thin or ruptured fibrous caps in advanced atherosclerosis. We investigated these associations in carotid arteries with mild to severe stenosis by in vivo 3T MRI. METHODS AND RESULTS: Seventy-seven patients with >or=50% carotid stenosis in at least one side by duplex ultrasound underwent bilateral multi-contrast carotid MRI scans. Measurements for wall and lipid-rich/necrotic core sizes, presence of hemorrhage, and fibrous cap status (classified as intact thick, intact thin or ruptured) were recorded. Arteries with poor image quality, occlusion, or no detectable lipid-rich/necrotic core were excluded. For the 798 MRI slices included, multivariate ordinal regression analysis demonstrated larger %lipid-rich/necrotic core (odds ratio for 10% increase, 1.49; P=0.02) and presence of hemorrhage (odds ratio, 5.91; P<0.001) were independently associated with a worse (intact thin or ruptured) stage of fibrous cap status. For artery-based multivariate analysis, a larger maximum %lipid-rich/necrotic core and presence of hemorrhage independently associated with worse fibrous cap status (P<0.001, for both). No hemorrhage was detected in arteries with thick fibrous caps. CONCLUSIONS: Hemorrhage and larger %lipid-rich/necrotic core were independently associated with a thin or ruptured fibrous cap status at an early to advanced stage of carotid atherosclerosis.

A study of images of Projective Angles of pulmonary veins.

AIMS: In images of magnetic resonance and computed tomography (CT) there are visible angles between pulmonary veins and the coronary, transversal or sagittal section of body. In this study these angles are measured and defined as Projective Angles of pulmonary veins. Several possible influential factors and characters of distribution are studied and analyzed for a better understanding of this imaging anatomic character of pulmonary veins. And it could be the anatomic base of adjusting correctly the angle of the central X-ray of the angiography of pulmonary veins undergoing the catheter ablation of atrial fibrillation (AF). METHOD: Images of contrast enhanced magnetic resonance angiography (CEMRA) and contrast enhanced computer tomography (CECT) of the left atrium and pulmonary veins of 137 health objects and patients with atrial fibrillation (AF) are processed with the technique of post-processing, and Projective Angles to the coronary and transversal sections are measured and analyzed statistically. RESULT: Project Angles of pulmonary veins are one of real and steady imaging anatomic characteristics of pulmonary veins. The statistical distribution of variables is relatively concentrated, with a fairly good representation of average value. It is possible to improve the angle of the central X-ray according to the average value in the selective angiography of pulmonary veins undergoing the catheter ablation of AF.