Engineering Yarrowia lipolytica for Sustainable Production of the Pomegranate Seed Oil-Derived Punicic Acid.

Punicic acid is a conjugated linolenic acid with various biological activities including antiobesity, antioxidant, anticancer, and anti-inflammatory effects. It is often used as a nutraceutical, dietary additive, and animal feed. Currently, punicic acid is primarily extracted from pomegranate seed oil, but it is restricted due to the extended growth cycle, climatic limitations, and low recovery level. There have also been reports on the chemical synthesis of punicic acid, but it resulted in a mixture of structurally similar isomers, requiring additional purification/separation steps. In this study, a comprehensive strategy for the production of punicic acid in Yarrowia lipolytica was implemented by pushing the supply of linoleic acid precursors in a high-oleic oil strain, expressing multiple copies of the fatty acid conjugase gene from Punica granatum, engineering the acyl-editing pathway to improve the phosphatidylcholine pool, and promoting the assembly of punicic acid in the form of triglycerides. The optimal strain with high oil production capacity and a significantly increased punicic acid ratio accumulated 3072.72 mg/L punicic acid, accounting for 6.19% of total fatty acids in fed-batch fermentation, providing a viable, sustainable, and green approach for punicic acid production to substitute plant extraction and chemical synthesis production.

High diagnostic performance of time-resolved MR angiography in spinal arteriovenous shunts.

BACKGROUND AND PURPOSE: In comparison with the limited efficacy of conventional MR imaging and the invasiveness of catheter-based digital subtraction angiography (DSA), time-resolved MR angiography (TR-MRA) has been proposed as a promising examination for early diagnosis of spinal arteriovenous shunts (SAVSs). This paper aims to investigate the diagnostic performance of TR-MRA with scan parameters optimized for SAVSs evaluation in a large number of patients. METHODS: One hundred patients with suspected SAVSs were enrolled. Each patient underwent preoperative TR-MRA with optimized scan parameters followed by DSA. The presence or absence of SAVS, the types and the angioarchitecture of SAVSs in the TR-MRA images were diagnostically analyzed. RESULTS: Among the final 97 patients, 80 cases (82.5 %) were diagnosed and classified by TR-MRA as spinal cord arteriovenous shunts (SCAVSs; n = 22), spinal dural arteriovenous shunts (SDAVSs; n = 48), and spinal extradural arteriovenous shunts (SEDAVSs; n = 10). The agreement for classifying SAVSs between TR-MRA and DSA was excellent (κ = 0.91). The sensitivity, specificity, PPV, NPV, and accuracy of TR-MRA for the diagnosis of SAVSs were 100 % (95 % CI, 94.3-100.0 %), 76.5 % (95 % CI, 49.8-92.2 %), 95.2 % (95 % CI, 87.6-98.5 %), 100 % (95 % CI, 71.7-100.0 %), and 95.9 % (95 % CI, 89.9-98.4 %). The accuracy rates of TR-MRA for the detection of feeding arteries were 75.9 %, 91.7 %, and 80.0 % for SCAVSs, SDAVSs, and SEDAVSs respectively. CONCLUSION: Time-resolved MR angiography showed excellent diagnostic performance for SAVSs screening. Additionally, this method can classify SAVSs and identify feeding arteries in SDAVSs with high diagnostic accuracy.

Age- and sex-specific reference values of biventricular strain and strain rate derived from a large cohort of healthy Chinese adults: a cardiovascular magnetic resonance feature tracking study.

BACKGROUND: As a noninvasive tool, myocardial deformation imaging may facilitate the early detection of cardiac dysfunction. However, normal reference ranges of myocardial strain and strain rate (SR) based on large-scale East Asian populations are still lacking. This study aimed to provide reference values of left ventricular (LV) and right ventricular (RV) strain and SR based on a large cohort of healthy Chinese adults using cardiovascular magnetic resonance (CMR) feature tracking (FT). METHODS: Five hundred and sixty-six healthy Chinese adults (55.1% men) free of hypertension, diabetes, and obesity were included. On cine CMR, biventricular global radial, circumferential, and longitudinal strain (GRS, GCS, and GLS), and the peak radial, circumferential, and longitudinal systolic, and diastolic SRs (PSSRR, PSSRC, PSSRL, PDSRR, PDSRC, and PDSRL), and regional radial and circumferential strain at the basal, mid-cavity, and apical levels were measured. Associations of global and regional biventricular deformation indices with age and sex were investigated. RESULTS: Women demonstrated greater magnitudes of LV GRS (37.6 ± 6.1% vs. 32.1 ± 5.3%), GCS (- 20.7 ± 1.9% vs. - 18.8 ± 1.9%), GLS (- 17.8 ± 1.8% vs. - 15.6 ± 1.8%), RV GRS (25.1 ± 7.8% vs. 22.1 ± 6.7%), GCS (- 14.4 ± 3.6% vs. - 13.2 ± 3.2%), GLS (- 22.4 ± 5.2% vs. - 20.2 ± 4.6%), and biventricular peak systolic and diastolic SR in all three coordinate directions (all P < 0.05). For the LV, aging was associated with increasing amplitudes of GRS, GCS, and decreasing amplitudes of PDSRR, PDSRC, PDSRL (all P < 0.05). For the RV, aging was associated with an increase in the magnitudes of GRS, GCS, GLS, PSSRR, PSSRC, PSSRL, and a decrease in the magnitude of PDSRR, PDSRC (all P < 0.05). Biventricular radial and circumferential strain measurements at the basal, mid-cavity, and apical levels were all significantly related to age and sex in both sexes (all P < 0.05). CONCLUSIONS: We provide age- and sex-specific normal values of biventricular strain and SR based on a large sample of healthy Chinese adults with a broad age range. These results may be served as a reference standard for cardiac function assessment, especially for the Chinese population.

Severe community-acquired pneumonia caused by Chlamydia pecorum.

Chlamydia pecorum is a zoonotic pathogen. Here, we report the first case of human infection with C. pecorum. A man aged 51 years with high fever and dry cough was diagnosed with severe community-acquired pneumonia and respiratory failure. C. pecorum was found responsible for the infection, which was detected from bronchoalveolar lavage fluid through metagenomic next-generation sequencing. C. pecorum infection was further identified by quantitative polymerase chain reaction and complement fixation test. The patient's condition improved rapidly after targeted treatment. He was a farmer with diabetes mellitus and had a history of close contact with sheep, which might result in C. pecorum infection. Our report could provide a direction for the diagnosis and treatment of human C. pecorum pneumonia.

Engineering Yarrowia lipolytica to produce nutritional fatty acids: Current status and future perspectives.

Due to their vital physiological functions, nutritional fatty acids have great potential as nutraceutical food supplements for preventing an array of diseases such as inflammation, depression, arthritis, osteoporosis, diabetes and cancer. Microbial biosynthesis of fatty acids follows the trend of sustainable development, as it enables green, environmentally friendly and efficient production. As a natural oleaginous yeast, Yarrowia lipolytica is especially well-suited for the production of fatty acids. Moreover, it has a variety of genetic engineering tools and novel metabolic engineering strategies that make it a robust workhorse for the production of an array of value-added products. In this review, we summarize recent advances in metabolic engineering strategies for accumulating nutritional fatty acids in Y. lipolytica, including conjugated fatty acids and polyunsaturated fatty acids. In addition, the future prospects of nutritional fatty acid production using the Y. lipolytica platform are discussed in light of the current progress, challenges, and trends in this field. Finally, guidelines for future studies are also emphasized.

Engineering Yarrowia lipolytica to produce advanced biofuels: Current status and perspectives.

Energy security and global climate change have necessitated the development of renewable energy with net-zero emissions. As alternatives to traditional fuels used in heavy-duty vehicles, advanced biofuels derived from fatty acids and terpenes have similar properties to current petroleum-based fuels, which makes them compatible with existing storage and transportation infrastructures. The fast development of metabolic engineering and synthetic biology has shown that microorganisms can be engineered to convert renewable feedstocks into these advanced biofuels. The oleaginous yeast Yarrowia lipolytica is rapidly emerging as a valuable chassis for the sustainable production of advanced biofuels derived from fatty acids and terpenes. Here, we provide a summary of the strategies developed in recent years for engineering Y. lipolytica to synthesize advanced biofuels. Finally, efficient biotechnological strategies for the production of these advanced biofuels and perspectives for future research are also discussed.

Biventricular Morphology and Function Reference Values Derived From a Large Sample of Healthy Chinese Adults by Magnetic Resonance Imaging.

Background: Quantification of cardiac structure and function is essential for diagnostic interpretation and clinical decision making. We sought to establish cardiovascular magnetic resonance (CMR) reference values of left and right ventricular (LV and RV) morphology and function based on a large sample of healthy Chinese adults. Methods: Five hundred fifty validated healthy Chinese adults (aged 21-70 years; 323 men) free of hypertension, diabetes, and obesity were included in this study. All the subjects were stratified by gender (men and women) and age decades. On cine CMR, measurements of biventricular end-diastolic, end-systolic, and stroke volumes (EDV, ESV, and SV), ejection fraction (EF), and end-diastolic LV wall thickness (LVWT) and mass (LVM) were obtained. Results: Men had greater LVEDV (111.6 ± 19.8 vs. 94.6 ± 15.6 ml), LVESV (36.5 ± 9.8 vs. 28.2 ± 7.9 ml), LVM (121.1 ± 19.9 vs. 86.1 ± 14.5 g), global end-diastolic LVWT (8.1 ± 1.1 vs. 6.7 ± 1.0 mm), RVEDV (128.0 ± 23.6 vs. 101.7 ± 17.0 ml), and RVESV (53.5 ± 13.7 vs. 36.8 ± 8.9 ml), while women had greater LVEF (67.5 ± 5.4 vs. 70.4 ± 5.7%) and RVEF (58.5 ± 5.2 vs. 64.0 ± 5.3%) (all p < 0.001). For both men and women, age was negatively correlated with LVEDV (r = -0.31 and r = -0.32), LVESV (r = -0.37 and r = -0.47), RVEDV (r = -0.31 and r = -0.29), and RVESV (r = -0.33 and r = -0.44), while it was positively correlated with LVEF (r = 0.28 and r = 0.43) and RVEF (r = 0.28 and r = 0.41) (all p < 0.001). Aging was associated with increasing global end-diastolic LVWT and LVM/LVEDV in both sexes (all p < 0.001). Older age was associated with increasing LVM only in women (r = 0.36, p < 0.001), not in men (r = 0.05, p = 0.359). Conclusions: We systematically provide age-, sex-, and body size-specific CMR reference values for biventricular morphology and function based on a large sample of healthy Chinese adults. Biventricular structure and function are significantly associated with age and sex.

Wall enhancement characteristics of vertebrobasilar nonsaccular aneurysms and their relationship to symptoms.

PURPOSE: This study aims to explore wall enhancement characteristics of vertebrobasilar nonsaccular aneurysms (VBNAs) and their relationship to clinical symptoms. METHODS: We retrospectively analyzed VBNAs in the high resolution MR vessel wall imaging (HR-VWI) databases at two institutions. The aneurysmal basic characteristics and wall enhancement characteristics, were evaluated by two radiologists. The aneurysmal basic characteristics were location, type, the maximal outer diameter (Dmax), T1 hyperintensity, and mural thrombus. The wall enhancement characteristics included the presence or absence of enhancement, enhancement distribution (concentric or eccentric), enhancement grades (<50% & 50%), enhancement index ((SIwall-post/SIbrain-post- SIwall-pre/SIbrain-pre)/ (SIwall-pre/SIbrain-pre)), enhancement ratio (SIwall-post/SIpituitary-post), and enhancement extent (focal BA, whole BA, and VBA). The VBNAs' related symptoms of all cases were identified. The associations of aneurysmal basic characteristics and wall enhancement characteristics with symptoms were analyzed. RESULTS: 34 VBNAs cases were included in the final analysis. Of the 34 cases, 18 cases (52.9%) presented VBNAs' related symptoms. Aneurysmal characteristics associated with symptoms were transitional type (P = 0.015), Dmax (P < 0.001), T1 hyperintensity (P = 0.003), mural thrombus (P < 0.001), the presence of enhancement (P = 0.003), enhancement index (P = 0.031), and enhancement ratio (P < 0.001). Aneurysmal characteristics that were independently associated with symptoms were Dmax (OR = 1.406, P = 0.013) and enhancement ratio (OR = 9.83, P = 0.037). In ROC curve analysis, AUC of the joint variable (Dmax and enhancement ratio) was 0.92, and the sensitivity and specificity were 94.4% and 87.5%, respectively. CONCLUSION: Wall enhancement correlates with symptomatic VBNAs. Dmax and enhancement ratio of aneurysmal wall are important indicators to instable status of symptomatic VBNAs.

Surveillance of abdominal aortic aneurysm using accelerated 3D non-contrast black-blood cardiovascular magnetic resonance with compressed sensing (CS-DANTE-SPACE).

BACKGROUND: 3D non-contrast high-resolution black-blood cardiovascular magnetic resonance (CMR) (DANTE-SPACE) has been used for surveillance of abdominal aortic aneurysm (AAA) and validated against computed tomography (CT) angiography. However, it requires a long scan time of more than 7 min. We sought to develop an accelerated sequence applying compressed sensing (CS-DANTE-SPACE) and validate it in AAA patients undergoing surveillance. METHODS: Thirty-eight AAA patients (all males, 73 ± 6 years) under clinical surveillance were recruited for this study. All patients were scanned with DANTE-SPACE (scan time 7:10 min) and CS-DANTE-SPACE (scan time 4:12 min, a reduction of 41.4%). Nine 9 patients were scanned more than 2 times. In total, 50 pairs of images were available for comparison. Two radiologists independently evaluated the image quality on a 1-4 scale, and measured the maximal diameter of AAA, the intra-luminal thrombus (ILT) and lumen area, ILT-to-muscle signal intensity ratio, and the ILT-to-lumen contrast ratio. The sharpness of the aneurysm inner/outer boundaries was quantified. RESULTS: CS-DANTE-SPACE achieved comparable image quality compared with DANTE-SPACE (3.15 ± 0.67 vs. 3.03 ± 0.64, p = 0.06). There was excellent agreement between results from the two sequences for diameter/area and ILT ratio measurements (ICCs> 0.85), and for quantifying growth rate (3.3 ± 3.1 vs. 3.3 ± 3.4 mm/year, ICC = 0.95.) CS-DANTE-SPACE showed a higher ILT-to-lumen contrast ratio (p = 0.01) and higher sharpness than DANTE-SPACE (p = 0.002). Both sequences had excellent inter-reader reproducibility for quantitative measurements (ICC > 0.88). CONCLUSION: CS-DANTE-SPACE can reduce scan time while maintaining image quality for AAA imaging. It is a promising tool for the surveillance of patients with AAA disease in the clinical setting.

Correlation between left ventricular myocardial strain and left ventricular geometry in healthy adults: a cardiovascular magnetic resonance-feature tracking study.

This study was aimed to investigate the correlation between left ventricular (LV) myocardial strain and LV geometry in healthy adults using cardiovascular magnetic resonance-feature tracking (CMR-FT). 124 gender-matched healthy adults who underwent healthy checkup using CMR cine imaging were retrospectively analyzed. Peak global radial, circumferential, longitudinal strain (GRS, GCS and GLS) for left ventricle were measured. LV geometry was assessed by the ratio of LV mass (LVM) and end-diastolic volume (EDV). GRS, GCS and GLS were 34.18 ± 6.71%, - 22.17 ± 2.28%, - 14.76 ± 2.39% for men, and 33.40 ± 6.95%, - 22.49 ± 2.27%, - 15.72 ± 2.36% for women. Multiple linear regression showed that LVM/EDV was associated with decreased GLS (ß = - 0.297, p = 0.005), but was not significantly associated with GRS and GCS (both p > 0.05). There was an increase in the magnitude of GRS, GCS and GLS with advancing age (ß = 0.254, ß = 0.466 and ß = 0.313, all p < 0.05). Greater BMI was associated with decreased GRS, GCS and GLS (ß = - 0.232, ß = - 0. 249 and ß = - 0.279, all p < 0.05). In conclusion, compared with GRS and GCS, GLS is more sensitive to assess LV concentric remodeling in healthy adults. GRS, GCS and GLS are all independently positively associated with age and negatively associated with BMI. Sex-based LV strain reference values for healthy Chinese adults are established.

Evaluation of the distribution and progression of intraluminal thrombus in abdominal aortic aneurysms using high-resolution MRI.

BACKGROUND: Intraluminal thrombus (ILT) signal intensity on MRI has been studied as a potential marker of abdominal aortic aneurysm (AAA) progression. PURPOSE: 1) To characterize the relationship between ILT signal intensity and AAA diameter; 2) to evaluate ILT change over time; and 3) to assess the relationship between ILT features and AAA growth. STUDY TYPE: Prospective. SUBJECTS: Eighty AAA patients were imaged, and a subset (n = 41) were followed with repeated MRI for 16 ± 9 months. FIELD STRENGTH/SEQUENCE: 3D black-blood fast-spin-echo sequence at 3 T. ASSESSMENT: ILT was designated as "bright" if the signal was greater than 1.2 times that of adjacent psoas muscle. AAAs were divided into three groups based on ILT: Type 1: bright ILT; Type 2: isointense ILT; Type 3: no ILT. During follow-up, an active ILT change was defined as new ILT formation or an increase in ILT signal intensity to bright; stable ILT was defined as no change in ILT type or ILT became isointense from bright previously. STATISTICAL TESTS: Shapiro-Wilk test; Mann-Whitney U-test; Fisher's exact test; Kruskal-Wallis test; Spearman's r; intraclass correlation coefficient (ICC), Cohen's kappa. RESULTS: AAAs with Type 1 ILT were larger than those with Types 2 and 3 ILT (5.1 ± 1.1 cm, 4.4 ± 0.9 cm, 4.2 ± 0.8 cm, P = 0.008). The growth rate of AAAs with Type 1 ILT was significantly greater than that of AAAs with Types 2 and 3 ILT (2.6 ± 2.5, 0.6 ± 1.3, 1.5 ± 0.6 mm/year, P = 0.01). During follow-up, AAAs with active ILT changes had a 3-fold increased growth rate compared with AAAs with stable ILT (3.6 ± 3.0 mm/year vs. 1.2 ± 1.5 mm/year, P = 0.008). DATA CONCLUSION: AAAs with bright ILT are larger in diameter and grow faster. Active ILT change is associated with faster AAA growth. Black-blood MRI can characterize ILT features and monitor their change over time, which may provide new insights into AAA risk assessment. LEVEL OF EVIDENCE: 2 Technical Efficacy Stage: 5 J. Magn. Reson. Imaging 2019;50:994-1001.

A distinct clinicopathological variant of focal cortical dysplasia IIId characterized by loss of layer 4 in the occipital lobe in 12 children with remote hypoxic-ischemic injury.

OBJECTIVE: In 2011, the International League Against Epilepsy (ILAE) proposed a consensus classification system of focal cortical dysplasia (FCD) to distinguish clinicopathological subtypes, for example, "isolated" FCD type Ia-c and IIa-b, versus "associated" FCD type IIIa-d. The histopathological differentiation of FCD type I and III variants remains, however, a challenging issue in everyday practice. We present a unique histopathological pattern in patients with difficult-to-diagnose FCD, which highlights this dilemma, but also helps to refine the current ILAE classification scheme of FCD. METHODS: We present a retrospective series of 11 male and one female patient with early onset pharmacoresistant epilepsy of the posterior quadrant (mean age at seizure onset = 4.6 years). All surgical specimens were reviewed. Clinical histories were retrieved and extracted from archival patient files. RESULTS: Microscopic inspection revealed abnormalities in cortical architecture with complete loss of layer 4 in all surgical samples of the occipital lobe, as confirmed by semiquantitative measurements (p < 0.01). Clinical history reported early transient hypoxic condition in nine patients (75%). Magnetic resonance imaging (MRI) revealed abnormal signals in the occipital lobe in all patients, and signal changes suggestive of subcortical encephalomalacia were found in seven patients. Surgical treatment achieved favorable seizure control (Engel class I and II) in seven patients with an available follow-up period of 6.1 years. SIGNIFICANCE: Prominent disorganization of cortical layering and lack of any other microscopically visible principle lesion in the surgical specimen would result in this neuropathological pattern hitherto being classified as FCD ILAE type Ib. However, perinatal hypoxia with distinctive MRI changes suggested primarily a hypoxemic lesion and acquired pathomechanism of neuronal cell loss in the occipital lobe of our patient series. We propose, therefore, classifying this distinctive clinicopathological pattern as a separate variant of FCD ILAE type IIId.

Prospective ECG-triggered coronary CT angiography: clinical value of noise-based tube current reduction method with iterative reconstruction.

OBJECTIVES: To evaluate the clinical value of noise-based tube current reduction method with iterative reconstruction for obtaining consistent image quality with dose optimization in prospective electrocardiogram (ECG)-triggered coronary CT angiography (CCTA). MATERIALS AND METHODS: We performed a prospective randomized study evaluating 338 patients undergoing CCTA with prospective ECG-triggering. Patients were randomly assigned to fixed tube current with filtered back projection (Group 1, n = 113), noise-based tube current with filtered back projection (Group 2, n = 109) or with iterative reconstruction (Group 3, n = 116). Tube voltage was fixed at 120 kV. Qualitative image quality was rated on a 5-point scale (1 = impaired, to 5 = excellent, with 3-5 defined as diagnostic). Image noise and signal intensity were measured; signal-to-noise ratio was calculated; radiation dose parameters were recorded. Statistical analyses included one-way analysis of variance, chi-square test, Kruskal-Wallis test and multivariable linear regression. RESULTS: Image noise was maintained at the target value of 35HU with small interquartile range for Group 2 (35.00-35.03HU) and Group 3 (34.99-35.02HU), while from 28.73 to 37.87HU for Group 1. All images in the three groups were acceptable for diagnosis. A relative 20% and 51% reduction in effective dose for Group 2 (2.9 mSv) and Group 3 (1.8 mSv) were achieved compared with Group 1 (3.7 mSv). After adjustment for scan characteristics, iterative reconstruction was associated with 26% reduction in effective dose. CONCLUSION: Noise-based tube current reduction method with iterative reconstruction maintains image noise precisely at the desired level and achieves consistent image quality. Meanwhile, effective dose can be reduced by more than 50%.

Noise-based tube current reduction method with iterative reconstruction for reduction of radiation exposure in coronary CT angiography.

PURPOSE: To investigate the potential of noise-based tube current reduction method with iterative reconstruction to reduce radiation exposure while achieving consistent image quality in coronary CT angiography (CCTA). MATERIALS AND METHODS: 294 patients underwent CCTA on a 64-detector row CT equipped with iterative reconstruction. 102 patients with fixed tube current were assigned to Group 1, which was used to establish noise-based tube current modulation formulas, where tube current was modulated by the noise of test bolus image. 192 patients with noise-based tube current were randomly assigned to Group 2 and Group 3. Filtered back projection was applied for Group 2 and iterative reconstruction for Group 3. Qualitative image quality was assessed with a 5 point score. Image noise, signal intensity, volume CT dose index, and dose-length product were measured. RESULTS: The noise-based tube current modulation formulas were established through regression analysis using image noise measurements in Group 1. Image noise was precisely maintained at the target value of 35.00 HU with small interquartile ranges for Group 2 (34.17-35.08 HU) and Group 3 (34.34-35.03 HU), while it was from 28.41 to 36.49 HU for Group 1. All images in the three groups were acceptable for diagnosis. A relative 14% and 41% reduction in effective dose for Group 2 and Group 3 were observed compared with Group 1. CONCLUSION: Adequate image quality could be maintained at a desired and consistent noise level with overall 14% dose reduction using noise-based tube current reduction method. The use of iterative reconstruction further achieved approximately 40% reduction in effective dose.

Evaluation of right ventricular function by 64-row CT in patients with chronic obstructive pulmonary disease and cor pulmonale.

PURPOSE: The aim of this study was to investigate the clinical application value of right ventricle (RV) function measured by 64 multi-detector row CT (MDCT) in patients with chronic obstructive pulmonary disease (COPD) and cor pulmonale. MATERIALS AND METHODS: Sixty-three consecutive patients with COPD and cor pulmonale were referred for electrocardiographically gated MDCT for evaluation of suspected or known coronary artery disease. Magnetic resonance imaging (MRI) for cardiac function analysis was performed on the same day. The MDCT and MRI examinations were successfully completed in 58 patients. Forty-six patients with COPD were divided into three groups according to the severity of disease by the pulmonary function test (PFT). Twelve patients diagnosed as cor pulmonale and 32 control subjects were also included. The RV function and myocardial mass (MM) were obtained by 64-MDCT and 1.5 T cardiac MRI in all of the groups. The results were compared among the groups using the Newman-Keuls method. Pearson's correlation was used to evaluate the relationship between the right ventricular ejection fraction (RVEF) and MM with the PFT results in COPD and cor pulmonale patients. RESULTS: The RVEF was significantly lower in patients with severe COPD and cor pulmonale than it was in those patients with mild or moderate COPD (P < 0.01). There were strong correlations between MDCT and MRI (r = 0.826 for RV MM, r = 0.982 and 0.969 for RV EDV and RV ESV, r = 0.899 for RVEF) and between MDCT results and forced expiratory volume in 1s (r = 0.787 for RVEF, r = -0.774 for RV MM) in all patients. CONCLUSION: MDCT can accurately quantify RV function and MM. The RVEF and RV MM measured by MDCT correlate well with the severity of disease as determined by PFT in patients with COPD and cor pulmonale. The assessment of right ventricular function is clinically important for evaluation of the severity of COPD, which may provide an objective basis for therapeutic strategy.

Optimization of contrast medium protocol in CT coronary arteriography: constant injection dose rate adapted to body weight.

BACKGROUND: Fixed dose contrast medium (CM) injection protocols are often used in CT coronary arteriography (CTCA). However, vascular enhancement is related to body weight and injection dose rate. PURPOSE: To compare aortic enhancement and coronary imaging quality between a fixed dose CM injection protocol and one adapted to body weight with a constant injection dose rate in mg I/kg/s. MATERIAL AND METHODS: One hundred consecutive patients undergoing CTCA were randomized to two groups; group I received 65 mL CM (370 mg I/mL) at 5 mL/s and group II received 0.9 mL CM (370 mg I/mL) per kg during 12 s resulting in a constant injection dose rate of 28 mg I/kg/s.The groups were re-divided according to body weight: IA ≤ 70 kg, IB >70 kg, IIA ≤ 70 kg, IIB >70 kg. Aortic attenuation was measured and coronary imaging quality was graded using a four-grade scale. RESULTS: The resulting mean CM dose, aortic attenuation and score of coronary imaging quality in groups IA/IB/IIA/IIB were 24/24/19/27 g I, 437/403/448/440 HU and 2.8/2.3/2.7/2.6, respectively. CM dose and aortic attenuation were significantly different between groups IA and IIA (p < 0.001, p = 0.01), as well as between groups IB and IIB (p < 0.001, p < 0.001).There was no statistically significant difference regarding coronary imaging quality between groups IA and IIA (p = 0.30), while there was a significant difference between groups IB and IIB (p < 0.001). CONCLUSION: In CTCA, body weight-adapted CM injection protocols using a constant dose rate can help save CM doses in slimmer patients and improve aortic enhancement and coronary imaging quality in heavier patients compared to fixed dose protocols.

Multiphase contrast-saline mixture injection with dual-flow in 64-row MDCT coronary CTA.

PURPOSE: To explore the feasibility of multiphase contrast-saline mixture with dual-flow injection technique for visualization of right ventricular (RV) cavity and interventricular septum (IVS) in 64-row multidetector computed tomography (MDCT) coronary angiography. MATERIALS AND METHODS: Twenty-four patients underwent coronary CT angiography (CTA) imaging with 64-row MDCT. In twelve patients (group A), 60 ml contrast medium (CM) bolus was followed by 40 ml saline, and in the other twelve patients (group B), 50 ml CM bolus was followed by 50 ml contrast-saline mixture at 60:40 ratio. The CM, saline and contrast-saline mixture flow rate were all 5.0 ml/s. Two experienced radiologists measured the CT values of ascending aorta, descending aorta, pulmonary artery and RV, rated the uniformity of RV cavity, the visualization of coronary arteries and IVS independently. RESULTS: By Kappa test, agreement between the two radiologists was 0.93 and 0.86 concerning the CT value measurements and the grades of the three indexes, respectively. By t-test, the mean CT values of ascending aorta and descending aorta of the two groups had no statistical difference (t=1.459, P>0.05; t=1.619, P>0.05); while the mean CT values of pulmonary artery and RV cavity had statistical differences (t=8.316, P<0.05; t=10.372, P<0.05). By two-related rank sum test, according to the visualization of coronary arteries and the uniformity of RV cavity, there were no statistical differences (U=66.00, P>0.05; U=54.00, P>0.05); while according to the visualization of IVS, group B was better than group A (U=8.00, P<0.05). CONCLUSION: In coronary CTA, a contrast-saline mixture after CM bolus can provide clear visualization of RV and IVS and LV without impairing coronary CTA image.

Role of Pax4 in Pdx1-VP16-mediated liver-to-endocrine pancreas transdifferentiation.

Although Pdx1-VP16 expression induces hepatic cell transdifferentiation into pancreatic precursor cells (WB-1), these incompletely reprogrammed cells fail to become glucose-sensitive insulin-producing cells in the absence of the activation of late-stage pancreatic transcription factors. As Pax4 promotes late-stage beta-cell differentiation and maturation, we generated lentiviral vector (LV) containing mouse Pax4 gene and developed two hepatic cell lines expressing Pax4 in the absence (WB-2 cells) or presence (WB-1A cells) of Pdx1-VP16, via LV-mediated gene transfer. Functional Pax4 protein expression in WB-2 and WB-1A cells was confirmed by electrophoretic mobility shift assay and Pdx1-VP16 protein expression in WB-1 and WB-1A cells was confirmed by Western blotting. Activation of Pax4 resulted in the expression of the late-stage transcription factors, including Pax6, Isl-1, and MafA, and generated a gene expression profile for WB-1A cells similar to that of functional rat insulinoma INS-1 cells. Insulin abundance in WB-1A cells was demonstrated by immunostaining. WB-1A cells exhibited glucose-responsive insulin release in vitro, and caused a rapid reversal of hyperglycemia following cell transplantation into streptozotocin-induced diabetic mice. Intraperitoneal glucose tolerance test showed a normal glucose response in WB-1, and WB-1A transplanted mice similar to that of normal mice. Removal of transplanted WB-1A cells resulted in a return of hyperglycemia, confirming that they were responsible for the observed normoglycemia. The explanted WB-1A cells exhibited strong insulin staining comparable to native islet beta-cells. These studies indicate that activation of Pax4 in Pdx1-VP16-expressing cells reprograms pancreatic precursor-like WB-1 cells into glucose-responsive, more mature insulin-producing cells.

Reprogramming liver-stem WB cells into functional insulin-producing cells by persistent expression of Pdx1- and Pdx1-VP16 mediated by lentiviral vectors.

Adenovirus-mediated transient expression of the pancreatic duodenal homeobox transcription factor Pdx1 in mouse liver activates pancreatic endocrine and exocrine genes, the latter reportedly resulting in severe hepatitis. Expression of a super-active form of Pdx1 or Pdx1-VP16 selectively transdifferentiates hepatic WB cells into functional pancreatic beta-like insulin-producing cells, without evidence of exocrine differentiation. No study has systematically compared the transdifferentiation efficiency of Pdx1 and Pdx1-VP16 at the cellular and molecular level. Comparisons can be ambiguous when vectors harboring a transcription factor cDNA have differing extents and duration of gene expression. In view of the remarkable capacity of lentiviral vector (LV) for delivering and integrating transgene into both dividing and nondividing cells, we transduced rat hepatic stem cell-like WB cells with LV-Pdx1 or LV-Pdx1-VP16, and then used the limiting-dilution technique to clone single-cell-derived cell lines that stably express either Pdx1 or Pdx1-VP16. With these cell lines, we studied: (a) the expression of Pdx1 or Pdx1-VP16 protein by Western blotting and immunocytochemistry; (b) the repertoire of long-term expression of Pdx1- or Pdx1-VP16-induced pancreatic gene expression using RT-PCR methods; and (c) their capacity to serve as beta-cell surrogates in restoring euglycemia in streptozotocin-treated diabetic mice. We found that cell lines expressing either Pdx1 or Pdx1-VP16 long-term exhibited similar profiles for expression of genes related to pancreatic development and beta-cell function, and reversed hyperglycemia in diabetic mice. We also examined short-term expression of Pdx1 or Pdx1-VP16, and the results demonstrated that expression of Pdx1-VP16 is more efficient in initiating liver-to-endocrine pancreas transdifferentiation. Our findings demonstrate: (a) that the LV system is highly effective in producing persistent expression of Pdx1 or Pdx1-VP16 in WB hepatic cells; and (b) long-term, persistent expression of either Pdx1 or Pdx1-VP16 is similarly effective in converting hepatic stem cells into pancreatic endocrine precursor cells that, upon transplantation into diabetic mice, become functional insulin-producing cells and restore euglycemia.

High glucose is necessary for complete maturation of Pdx1-VP16-expressing hepatic cells into functional insulin-producing cells.

Pdx1 has been shown to convert hepatocytes into both exocrine and endocrine pancreatic cells in mice, but it fails to selectively convert hepatocytes into pure insulin-producing cells (IPCs). The molecular mechanisms underlying the transdifferentiation remain unclear. In this study, we generated a stably transfected rat hepatic cell line named WB-1 that expresses an active form of Pdx1 along with a reporter gene, RIP-eGFP. Our results demonstrate that Pdx1 induces the expression of multiple genes related to endocrine pancreas development and islet function in these liver cells. We do not however find any expression of the late-stage genes (Pax4, Pax6, Isl-1, and MafA) related to beta-cell development, and the cells do not secrete insulin upon the glucose challenge. Yet when WB-1 cells are transplanted into diabetic NOD-scid mice, these genes become activated and hyperglycemia is completely reversed. Detailed comparison of gene expression profiles between pre- and posttransplanted WB-1 cells demonstrates that the WB-1 cells have similar properties as that seen in pancreatic beta-cells. In addition, in vitro culture in high-glucose medium is sufficient to induce complete maturation of WB-1 cells into functional IPCs. In summary, we find that Pdx1-VP16 is able to selectively convert hepatic cells into pancreatic endocrine precursor cells. However, complete transdifferentiation into functional IPCs requires additional external factors, including high glucose or hyperglycemia. Thus, transdifferentiation of hepatocytes into functional IPCs may serve as a viable therapeutic option for patients with type 1 diabetes.