Chloroplast SRP54 and FtsH protease coordinate thylakoid membrane-associated proteostasis in Arabidopsis.

Thylakoid membrane protein quality control (PQC), which requires the coordination of membrane protein translocation and degradation of unassembled proteins, determines chloroplast development during de-etiolation. Despite numerous efforts, the regulation of this process in land plants is largely unknown. Here, we report the isolation and characterization of pale green Arabidopsis4 (pga4) mutants in Arabidopsis (Arabidopsis thaliana) with defects in chloroplast development during de-etiolation. Map-based cloning and complementation assays confirmed that PGA4 encodes the chloroplast Signal Recognition Particle 54 kDa (cpSRP54) protein. A heterogeneous Light-Harvesting Chlorophyll a/b Binding-Green Fluorescent Protein (LhcB2-GFP) fusion protein was generated as an indicative reporter for cpSRP54-mediated thylakoid translocation. LhcB2-GFP was dysfunctional and degraded to a short-form dLhcB2-GFP during de-etiolation through an N-terminal degradation initiated on thylakoid membranes. Further biochemical and genetic evidence demonstrated that the degradation of LhcB2-GFP to dLhcB2-GFP was disrupted in pga4 and yellow variegated2 (var2) mutants caused by mutations in the Filamentous Temperature-Sensitive H2 (VAR2/AtFtsH2) subunit of thylakoid FtsH. The yeast two-hybrid assay showed that the N-terminus of LhcB2-GFP interacts with the protease domain of VAR2/AtFtsH2. Moreover, the over-accumulated LhcB2-GFP in pga4 and var2 formed protein aggregates, which were insoluble in mild nonionic detergents. Genetically, cpSRP54 is a suppressor locus for the leaf variegation phenotype of var2. Together, these results demonstrate the coordination of cpSRP54 and thylakoid FtsH in maintaining thylakoid membrane PQC during the assembly of photosynthetic complexes and provide a trackable substrate and product for monitoring cpSRP54-dependent protein translocation and FtsH-dependent protein degradation.

Chloroplast envelope ATPase PGA1/AtFtsH12 is required for chloroplast protein accumulation and cytosol-chloroplast protein homeostasis in Arabidopsis.

The establishment of photosynthetic protein complexes during chloroplast development requires the influx of a large number of chloroplast proteins that are encoded by the nuclear genome, which is critical for cytosol and chloroplast protein homeostasis and chloroplast development. However, the mechanisms regulating this process are still not well understood in higher plants. Here, we report the isolation and characterization of the pale green Arabidopsis pga1-1 mutant, which is defective in chloroplast development and chloroplast protein accumulation. Using genetic and biochemical evidence, we reveal that PGA1 encodes AtFtsH12, a chloroplast envelope-localized protein of the FtsH family proteins. We determined a G703R mutation in the GAD motif of the conserved ATPase domain renders the pga1-1 a viable hypomorphic allele of the essential gene AtFtsH12. In de-etiolation assays, we showed that the accumulation of photosynthetic proteins and the expression of photosynthetic genes were impaired in pga1-1. Using the FNRctp-GFP and pTAC2-GFP reporters, we demonstrated that AtFtsH12 was required for the accumulation of chloroplast proteins in vivo. Interestingly, we identified an increase in expression of the mutant AtFtsH12 gene in pga1-1, suggesting a feedback regulation. Moreover, we found that cytosolic and chloroplast proteostasis responses were triggered in pga1-1. Together, taking advantage of the novel pga1-1 mutant, we demonstrate the function of AtFtsH12 in chloroplast protein homeostasis and chloroplast development.

Deep Learning for Fully Automated Prediction of Overall Survival in Patients Undergoing Resection for Pancreatic Cancer: A Retrospective Multicenter Study.

OBJECTIVE: To develop an imaging-derived biomarker for prediction of overall survival (OS) of pancreatic cancer by analyzing preoperative multiphase contrast-enhanced computed topography (CECT) using deep learning. BACKGROUND: Exploiting prognostic biomarkers for guiding neoadjuvant and adjuvant treatment decisions may potentially improve outcomes in patients with resectable pancreatic cancer. METHODS: This multicenter, retrospective study included 1516 patients with resected pancreatic ductal adenocarcinoma (PDAC) from 5 centers located in China. The discovery cohort (n=763), which included preoperative multiphase CECT scans and OS data from 2 centers, was used to construct a fully automated imaging-derived prognostic biomarker-DeepCT-PDAC-by training scalable deep segmentation and prognostic models (via self-learning) to comprehensively model the tumor-anatomy spatial relations and their appearance dynamics in multiphase CECT for OS prediction. The marker was independently tested using internal (n=574) and external validation cohorts (n=179, 3 centers) to evaluate its performance, robustness, and clinical usefulness. RESULTS: Preoperatively, DeepCT-PDAC was the strongest predictor of OS in both internal and external validation cohorts [hazard ratio (HR) for high versus low risk 2.03, 95% confidence interval (CI): 1.50-2.75; HR: 2.47, CI: 1.35-4.53] in a multivariable analysis. Postoperatively, DeepCT-PDAC remained significant in both cohorts (HR: 2.49, CI: 1.89-3.28; HR: 2.15, CI: 1.14-4.05) after adjustment for potential confounders. For margin-negative patients, adjuvant chemoradiotherapy was associated with improved OS in the subgroup with DeepCT-PDAC low risk (HR: 0.35, CI: 0.19-0.64), but did not affect OS in the subgroup with high risk. CONCLUSIONS: Deep learning-based CT imaging-derived biomarker enabled the objective and unbiased OS prediction for patients with resectable PDAC. This marker is applicable across hospitals, imaging protocols, and treatments, and has the potential to tailor neoadjuvant and adjuvant treatments at the individual level.

Interaction of PALE CRESS with PAP2/pTAC2 and PAP3/pTAC10 affects the accumulation of plastid-encoded RNA polymerase complexes in Arabidopsis.

The transcription of photosynthesis genes in chloroplasts is largely mediated by the plastid-encoded RNA polymerase (PEP), which resembles prokaryotic-type RNA polymerases, but with plant-specific accessory subunits known as plastid transcriptionally active chromosome proteins (pTACs) or PEP-associated proteins (PAPs). However, whether additional factors are involved in the biogenesis of PEP complexes remains unknown. Here, we investigated the function of an essential gene, PALE CRESS (PAC), in the accumulation of PEP complexes in chloroplasts. We established that an Arabidopsis leaf variegation mutant, variegated 6-1 (var6-1), is a hypomorphic allele of PAC. Unexpectedly, we revealed that a fraction of VAR6/PAC is associated with thylakoid membranes, where it interacts with PEP complexes. The accumulation of PEP complexes is defective in both var6-1 and the null allele var6-2. Further protein interaction assays confirmed that VAR6/PAC interacts directly with the PAP2/pTAC2 and PAP3/pTAC10 subunits of PEP complexes. Moreover, we generated viable hypomorphic alleles of the essential gene PAP2/pTAC2, and revealed a genetic interaction between PAC and PAP2/pTAC2 in photosynthesis gene expression and PEP complex accumulation. Our findings establish that VAR6/PAC affects PEP complex accumulation through interactions with PAP2/pTAC2 and PAP3/pTAC10, and provide new insights into the accumulation of PEP and chloroplast development.

PD-L1 Aptamer-Functionalized Metal-Organic Framework Nanoparticles for Robust Photo-Immunotherapy against Cancer with Enhanced Safety.

Immune checkpoint blockade has become a paradigm-shifting treatment modality to combat cancer, while conventional administration of immune checkpoint inhibitors, such as anti-PD-L1 antibody (α-PD-L1), often shows unsatisfactory immune responses and lead to severe immune-related adverse effects (irAEs). Herein, we develop a PD-L1 aptamer-based spherical nucleic acids (SNAs), which consists of oxaliplatin (OXA) encapsulated in a metal-organic framework nanoparticle core and a dense shell of aptPD-L1 (denoted as M@O-A). Upon light irradiation, this nanosystem enables concurrent photodynamic therapy (PDT), chemotherapy, and enhanced immunotherapy in one shot to inhibit both primary colorectal tumors and untreated distant tumors in mice. Notably, M@O-A shows scarcely any systemic immunotoxicity in a clinical irAEs-mimic transgenic mouse model. Collectively, this study presents a novel strategy for priming robust photo-immunotherapy against cancer with enhanced safety.

VAR2/AtFtsH2 and EVR2/BCM1/CBD1 synergistically regulate the accumulation of PSII reaction centre D1 protein during de-etiolation in Arabidopsis.

Thylakoid FtsH complex participates in PSII repair cycle during high light-induced photoinhibition. The Arabidopsis yellow variegated2 (var2) mutants are defective in the VAR2/AtFtsH2 subunit of thylakoid FtsH complex. Taking advantage of the var2 leaf variegation phenotype, dissections of genetic enhancer loci have yielded novel paradigms in understanding functions of thylakoid FtsH complex. Here, we report the isolation of a new var2 enhancer, enhancer of variegation2-1 (evr2-1). We confirmed that EVR2 encodes a chloroplast protein that was known as BALANCE OF CHLOROPHYLL METABOLISM 1 (BCM1), or CHLOROPHYLL BIOSYNTHETIC DEFECT 1 (CBD1). We showed that EVR2/BCM1/CBD1 was involved in the oligomerization of photosystem I complexes. Genetic assays indicated that general defects in chlorophyll biosynthesis and the accumulation of photosynthetic complexes do not necessarily enhance var2 leaf variegation. In addition, we found that VAR2/AtFtsH2 is required for the accumulation of photosynthetic proteins during de-etiolation. Moreover, we identified PSII core proteins D1 and PsbC as potential EVR2-associated proteins using Co-IP/MS. Furthermore, the accumulation of D1 protein was greatly compromised in the var2-5 evr2-1 double mutant during de-etiolation. Together, our findings reveal a functional link between VAR2/AtFtsH2 and EVR2/BCM1/CBD1 in regulating chloroplast development and the accumulation of PSII reaction centre D1 protein during de-etiolation.

MRI-derived radiomics analysis improves the noninvasive pretreatment identification of multimodality therapy candidates with early-stage cervical cancer.

OBJECTIVES: To develop and validate a clinical-radiomics model that incorporates radiomics signatures and pretreatment clinicopathological parameters to identify multimodality therapy candidates among patients with early-stage cervical cancer. METHODS: Between January 2017 and February 2021, 235 patients with IB1-IIA1 cervical cancer who underwent radical hysterectomy were enrolled and divided into training (n = 194, training:validation = 8:2) and testing (n = 41) sets according to surgical time. The radiomics features of each patient were extracted from preoperative sagittal T2-weighted images. Significance testing, Pearson correlation analysis, and Least Absolute Shrinkage and Selection Operator were used to select radiomic features associated with multimodality therapy administration. A clinical-radiomics model incorporating radiomics signature, age, 2018 Federation International of Gynecology and Obstetrics (FIGO) stage, menopausal status, and preoperative biopsy histological type was developed to identify multimodality therapy candidates. A clinical model and a clinical-conventional radiological model were also constructed. A nomogram and decision curve analysis were developed to facilitate clinical application. RESULTS: The clinical-radiomics model showed good predictive performance, with an area under the curve, sensitivity, and specificity in the testing set of 0.885 (95% confidence interval: 0.781-0.989), 78.9%, and 81.8%, respectively. The AUC, sensitivity, and specificity of the clinical model and clinical-conventional radiological model were 0.751 (0.603-0.900), 63.2%, and 63.6%, 0.801 (0.661-0.942), 73.7%, and 68.2%, respectively. A decision curve analysis demonstrated that when the threshold probability was > 20%, the clinical-radiomics model or nomogram may be more advantageous than the treat all or treat-none strategy. CONCLUSIONS: The clinical-radiomics model and nomogram can potentially identify multimodality therapy candidates in patients with early-stage cervical cancer. KEY POINTS: • Pretreatment identification of multimodality therapy candidates among patients with early-stage cervical cancer helped to select the optimal primary treatment and reduce severe complication risk and costs. • The clinical-radiomics model achieved a better prediction performance compared with the clinical model and the clinical-conventional radiological model. • An easy-to-use nomogram exhibited good performance for individual preoperative prediction.

Assessment of Changes in the Cesarean Scar and Uterus Between One and Two Years after Cesarean Section Using 3D T2w SPACE MRI.

Objective To evaluate changes in morphology of the cesarean scar and uterus between one and two years after cesarean section using high-resolution, three dimensional T2-weighted sampling perfection with application optimized contrast using different flip angle evolutions Magnetic Resonance Imaging (3D T2w SPACE MRI). Methods This prospective study was performed to investigate morphological changes in the cesarean scars and uterus from one to two years after cesarean section using high-resolution, 3D T2w SPACE MRI. The healthy volunteers having no childbearing history were recruited as the controls. All data were measured by two experienced radiologists. All data with normal distribution between the one-year and two-year groups were compared using a paired-sample t test or independent t test. Results Finally, 46 women took a pelvic MR examination one year after cesarean section, and a subset of 15 completed the same examination again after two years of cesarean section. Both the uterine length and the anterior wall thickness after two years of cesarean section (5.75 ± 0.46 and 1.45 ± 0.35 cm) were significantly greater than those measured at one year (5.33 ± 0.59 and 1.25 ± 0.27 cm) (t = -2.363 and -2.175, P= 0.033 and 0.048). No significant difference was shown in myometrial thickness two years after cesarean section (1.45 ±0.35 cm) with respect to the control group (1.58 ± 0.21 cm, P= 0.170). Nine women who underwent MRI twice were considered to have scar diverticula one year after cesarean section, and still had diverticula two years after cesarean section. The thickness, height, and width of the uterine scar showed no significant change from one to two years (all P > 0.05). Conclusions 3D T2w SPACE MRI provides overall morphologic details and shows dynamic changes in the scar and the uterus between one and two years after cesarean section. Scar morphology after cesarean section reached relatively stable one year after cesarean section, and uterine morphology was closer to normal two years after cesarean section.

The chloroplast metalloproteases VAR2 and EGY1 act synergistically to regulate chloroplast development in Arabidopsis.

Chloroplast development and photosynthesis require the proper assembly and turnover of photosynthetic protein complexes. Chloroplasts harbor a repertoire of proteases to facilitate proteostasis and development. We have previously used an Arabidopsis leaf variegation mutant, yellow variegated2 (var2), defective in thylakoid FtsH protease complexes, as a tool to dissect the genetic regulation of chloroplast development. Here, we report a new genetic enhancer mutant of var2, enhancer of variegation3-1 (evr3-1). We confirm that EVR3 encodes a chloroplast metalloprotease, reported previously as ethylene-dependent gravitropism-deficient and yellow-green1 (EGY1)/ammonium overly sensitive1 (AMOS1). We observed that mutations in EVR3/EGY1/AMOS1 cause more severe leaf variegation in var2-5 and synthetic lethality in var2-4 Using a modified blue-native PAGE system, we reveal abnormal accumulations of photosystem I, photosystem II, and light-harvesting antenna complexes in EVR3/EGY1/AMOS1 mutants. Moreover, we discover distinct roles of VAR2 and EVR3/EGY1/AMOS1 in the turnover of photosystem II reaction center under high light stress. In summary, our findings indicate that two chloroplast metalloproteases, VAR2/AtFtsH2 and EVR3/EGY1/AMOS1, function coordinately to regulate chloroplast development and reveal new roles of EVR3/EGY1/AMOS1 in regulating chloroplast proteostasis in Arabidopsis.

Three-dimensional MR Elastography Depicts Liver Inflammation, Fibrosis, and Portal Hypertension in Chronic Hepatitis B or C.

Background The value of measuring mechanical properties to categorize various pathophysiologic states of the liver is as yet undetermined in chronic hepatitis B (CHB) or C (CHC). Purpose To evaluate multiparametric three-dimensional (3D) MR elastography as a means of detecting early necroinflammation, distinguishing necroinflammation from fibrosis, and gauging the severity of portal hypertension (PH) in CHB or CHC. Materials and Methods From January 2015 to September 2019, participants with CHB or CHC were prospectively enrolled from a single institution and were divided into two groups: those with liver biopsy and no evidence of PH (group 1) and those with PH and a hepatic venous pressure gradient (HVPG) measurement (group 2). For group 3, healthy volunteers were separately recruited from a nearby community. Multiple viscoelastic parameters (shear stiffness [SS], storage modulus, loss modulus, and damping ratio [DR]) were determined at 3D MR elastography at 60 Hz, and multivariable logistic or linear regression analysis was used to assess associations of mechanical parameters with histologic scores and HVPG. Results A total of 155 participants (median age, 41 years [interquartile range, 32-48 years]; 85 women) were in group 1 (training set: n = 78, validation set: n = 77), 85 participants (median age, 57 years [interquartile range, 43-61 years]; 51 women) in group 2, and 60 healthy volunteers (median age, 49 years [interquartile range, 27-64 years]; 38 men) in group 3. The liver DR was higher in participants with necroinflammation (DR, 0.13 ± 0.03) versus those without (at liver fibrosis stage F0) (DR, 0.10 ± 0.02; P < .001). Liver DR and SS together performed well in the diagnosis of necroinflammation (area under the receiver operating characteristic curve [AUC], 0.88 [95% CI: 0.79, 0.96]) and the scoring of moderate to severe activity (AUC, 0.88 [95% CI: 0.81, 0.95]) in the validation data set. Liver DR (regression coefficient [ß] = -30.3 [95% CI: -58.0, -2.5]; P = .03) and splenic SS (ß = 2.3 [95% CI: 1.7, 2.9]; P < .001) were independently associated with HVPG. Conclusion Three-dimensional MR elastography may detect early necroinflammation, distinguish necroinflammation from liver fibrosis, and correlate with hepatic venous pressure gradient in chronic hepatitis B and C. © RSNA, 2021 Online supplemental material is available for this article. See also the editorial by Reeder in this issue.

NGATHA-LIKEs Control Leaf Margin Development by Repressing CUP-SHAPED COTYLEDON2 Transcription.

The leaf margin is a fascinating feature of leaf morphology, contributing to the incredible diversity of leaf shapes and forms. As a central regulator of plant organ separation and margin development, CUP-SHAPED COTYLEDON2 (CUC2), a NAM, ATAF1, 2, CUC2 (NAC)-family transcription factor, governs the extent of serrations along the leaf margin. CUC2 activity is tightly regulated at transcriptional and posttranscriptional levels. However, the molecular mechanism that controls CUC2 transcription during leaf development has not been fully elucidated. Here we report that Arabidopsis (Arabidopsis thaliana) NGATHA-LIKE1 (NGAL1) to NGAL3, which are three related B3 family transcription factors, act as negative regulators of leaf margin serration formation. Over-expression of NGALs led to "cup-shaped" cotyledons and smooth leaf margins, whereas the triple loss-of-function mutant ngaltri exhibited more serrated leaves than the wild type. RNA-sequencing analyses revealed that the expression levels of a number of transcription factor genes involved in leaf development are regulated by NGALs, including CUC2 Comparative transcriptome analyses further uncovered a significant overlap between NGAL- and CUC2-regulated genes. Moreover, genetic analyses using various combinations of gain- and loss-of-function mutants of NGALs and CUC2 confirmed that CUC2 acts downstream of NGALs in promoting the formation of leaf-margin serrations. Finally, we demonstrate that NGAL1 directly binds to the CUC2 promoter causing repressed CUC2 expression. In summary, direct CUC2 transcriptional repression by NGAL1 characterizes a further regulatory module controlling leaf margin development.

ABNORMAL SHOOT 6 interacts with KATANIN 1 and SHADE AVOIDANCE 4 to promote cortical microtubule severing and ordering in Arabidopsis.

Plant interphase cortical microtubules (cMTs) mediate anisotropic cell expansion in response to environmental and developmental cues. In Arabidopsis thaliana, KATANIN 1 (KTN1), the p60 catalytic subunit of the conserved MT-severing enzyme katanin, is essential for cMT ordering and anisotropic cell expansion. However, the regulation of KTN1-mediated cMT severing and ordering remains unclear. In this work, we report that the Arabidopsis IQ67 DOMAIN (IQD) family gene ABNORMAL SHOOT 6 (ABS6) encodes a MT-associated protein. Overexpression of ABS6 leads to elongated cotyledons, directional pavement cell expansion, and highly ordered transverse cMT arrays. Genetic suppressor analysis revealed that ABS6-mediated cMT ordering is dependent on KTN1 and SHADE AVOIDANCE 4 (SAV4). Live imaging of cMT dynamics showed that both ABS6 and SAV4 function as positive regulators of cMT severing. Furthermore, ABS6 directly interacts with KTN1 and SAV4 and promotes their recruitment to the cMTs. Finally, analysis of loss-of-function mutant combinations showed that ABS6, SAV4, and KTN1 work together to ensure the robust ethylene response in the apical hook of dark-grown seedlings. Together, our findings establish ABS6 and SAV4 as positive regulators of cMT severing and ordering, and highlight the role of cMT dynamics in fine-tuning differential growth in plants.

LncRNA NKILA inhibits the proliferation and promotes the apoptosis of CSCC cells by downregulating miRNA-21.

Long noncoding RNA (lncRNA) NKILA has been well studied in several types of human tumors as a tumor suppressor, while its involvement in cervical squamous cell carcinoma (CSCC) remains unclear. In our studies, we found that serum NKILA was at lower levels and serum microRNA-21 (miRNA-21) was at higher levels in patients with early stage CSCC than in the healthy female. Altered expression of NKILA and miRNA-21 can effectively separate patients with CSCC at an early stage from healthy controls. Serum levels of NKILA were significantly and negatively correlated with miRNA-21 in patients with CSCC but not in normal controls. Overexpression of NKILA mediated the inhibited expression of miRNA-21 in CSCC cells, but mimic transfection of miRNA-21 did not significantly change the expression level of NKILA. Overexpression of NKILA repressed the proliferation and promoted the apoptosis of CSCC cells, while miRNA-21 showed opposite functions. In addition, miRNA-21 mimic transfection reduced the effects of NKILA on CSCC cells. Collectively, lncRNA NKILA could repress the proliferation and promote the apoptosis of CSCC cells by downregulating miRNA-21.

Multiparametric MRI-Based Radiomics for Prostate Cancer Screening With PSA in 4-10 ng/mL to Reduce Unnecessary Biopsies.

BACKGROUND: Whether men with a prostate-specific antigen (PSA) level of 4-10 ng/mL should be recommended for a biopsy is clinically challenging. PURPOSE: To develop and validate a radiomics model based on multiparametric MRI (mp-MRI) in patients with PSA levels of 4-10 ng/mL to predict prostate cancer (PCa) preoperatively and reduce unnecessary biopsies. STUDY TYPE: Retrospective. SUBJECTS: In all, 199 patients with PSA levels of 4-10 ng/mL. FIELD STRENGTH/SEQUENCE: 3T, T2 -weighted, diffusion-weighted, and dynamic contrast-enhanced MRI. ASSESSMENT: Lesion regions of interest (ROIs) from T2 -weighted, diffusion-weighted, and dynamic contrast-enhanced MRI were annotated by two radiologists. A total of 2104 radiomic features were extracted from the ROI of each patient. A random forest classifier was used to build the radiomics model for PCa in the primary cohort. A combined model was constructed using multivariate logistic regression by incorporating the radiomics signature and clinical-radiological risk factors. STATISTICAL TESTS: For continuous variables, variance equality was assessed by Levene's test and Student's t-test, and Welch's t-test was used to assess between-group differences. For categorical variables, Pearson's chi-square test, Fisher's exact test, or the approximate chi-square test was used to assess between-group differences. P < 0.05 was considered statistically significant. RESULTS: The combined model incorporating the multi-imaging fusion model, age, PSA density (PSAD), and the PI-RADS v2 score yielded area under the curve (AUC) values of 0.956 and 0.933 on the primary (n = 133) and validation (n = 66) cohorts, respectively. Compared with the clinical-radiological model, the combined model performed better on both the primary and validation cohorts (P < 0.05). Furthermore, the use of the combined model to predict PCa could identify more negative PCa patients than the use of the clinical-radiological model by 18.4%. DATA CONCLUSION: The combined model was developed and validated to provide potential preoperative prediction of PCa in men with PSA levels of 4-10 ng/mL and might aid in treatment decision-making and reduce unnecessary biopsies. LEVEL OF EVIDENCE: 3 Technical Efficacy Stage: 3 J. Magn. Reson. Imaging 2020;51:1890-1899.

Radiomics Based on MRI as a Biomarker to Guide Therapy by Predicting Upgrading of Prostate Cancer From Biopsy to Radical Prostatectomy.

BACKGROUND: Biopsy Gleason score (GS) is crucial for prostate cancer (PCa) treatment decision-making. Upgrading in GS from biopsy to radical prostatectomy (RP) puts a proportion of patients at risk of undertreatment. PURPOSE: To develop and validate a radiomics model based on multiparametric magnetic resonance imaging (mp-MRI) to predict PCa upgrading. STUDY TYPE: Retrospective, radiomics. POPULATION: A total of 166 RP-confirmed PCa patients (training cohort, n = 116; validation cohort, n = 50) were included. FIELD STRENGTH/SEQUENCE: 3.0T/T2 -weighted (T2 W), apparent diffusion coefficient (ADC), and dynamic contrast enhancement (DCE) sequences. ASSESSMENT: PI-RADSv2 score for each tumor was recorded. Radiomic features were extracted from T2 W, ADC, and DCE sequences and Mutual Information Maximization criterion was used to identify the optimal features on each sequence. Multivariate logistic regression analysis was used to develop predictive models and a radiomics nomogram and their performance was evaluated. STATISTICAL TESTS: Student's t or chi-square were used to assess the differences in clinicopathologic data between the training and validation cohorts. Receiver operating characteristic (ROC) curve analysis was performed and the area under the curve (AUC) was calculated. RESULTS: In PI-RADSv2 assessment, 67 lesions scored 5, 70 lesions scored 4, and 29 lesions scored 3. For each sequence, 4404 features were extracted and the top 20 best features were selected. The radiomics model incorporating signatures from the three sequences achieved better performance than any single sequence (AUC: radiomics model 0.868, T2 W 0.700, ADC 0.759, DCE 0.726). The combined mode incorporating radiomics signature, clinical stage, and time from biopsy to RP outperformed the clinical model and radiomics model (AUC: combined model 0.910, clinical model 0.646, radiomics model 0.868). The nomogram showed good performance (AUC 0.910) and calibration (P-values: training cohort 0.624, validation cohort 0.294). DATA CONCLUSION: Radiomics based on mp-MRI has potential to predict upgrading of PCa from biopsy to RP. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 5 J. Magn. Reson. Imaging 2020;52:1239-1248.

Pancreatic adenocarcinoma: quantitative CT features are correlated with fibrous stromal fraction and help predict outcome after resection.

OBJECTIVES: To identify quantitative imaging features of contrast-enhanced computed tomography (CE-CT) that may be prognostically favorable after resection of smaller (≤ 30 mm) pancreatic ductal adenocarcinomas (PDACs) located at head. METHODS: This retrospective study included two independent cohorts (discovery cohort, n = 212; test cohort, n = 100) of patients who underwent resection of head PDACs ≤ 30 mm and preoperative CE-CT. We examined tumor and surrounding parenchymal attenuation differences (deltas), and tumor attenuation changes across phases (ratios). Semantic features of PDACs were evaluated by two radiologists. Clinicopathologic and imaging features for predicting disease-free survival (DFS) and overall survival (OS) were analyzed via multivariate Lasso-penalized Cox proportional-hazards models. Survival rates were derived by Kaplan-Meier method. RESULTS: Imaging features achieved C-indices of 0.766 (discovery cohort) and 0.739 (test cohort) for DFS, and 0.790 (discovery cohort) and 0.772 (test cohort) for OS estimates through incorporation of clinicopathologic features. The most decisive imaging feature was delta 3, denoting attenuation differences between tumor and surrounding pancreas at pancreatic phase (DFS: HR = 2.122; OS: HR = 2.375; both p < 0.001). Compared with inconspicuous (low-delta-3, < 28 HU) tumors, conspicuous (high-delta-3) tumors correlated significantly with more aggressive histologic grades (p = 0.014) and less extensive tumor fibrous stromal fractions (p < 0.001). Patients with low-delta-3 tumors ≤ 20 mm experienced the most favorable outcomes (DFS, 36 months; OS, 42 months), whereas those with high-delta-3 tumors fared poorly, regardless of tumor size (DFS, 12 months; OS, 19 months). CONCLUSIONS: Quantifiable CT imaging features reflect heterogeneous fibrous stromal fractions and histologic grades of PDAC at head locations that help stratify patients with disparate clinical outcomes. KEY POINTS: • Quantitative and semantic imaging features achieved promising results for the prognosis of resected PDAC (≤ 30 mm) at head location, through incorporation of clinicopathologic features. • Attenuation difference at tumor-parenchyma interface (delta 3) emerged as the most decisive imaging feature, enabling further stratification of patients into distinct prognostic subtypes by tumor size. • High delta 3 signifies sharper contrast between tumor and surrounding pancreas, correlating with more aggressive histologic grades and less extensive tumor fibrous stromal fractions.

CT radiomics nomogram for the preoperative prediction of lymph node metastasis in gastric cancer.

PURPOSE: To investigate the role of computed tomography (CT) radiomics for the preoperative prediction of lymph node (LN) metastasis in gastric cancer. MATERIALS AND METHODS: This retrospective study included 247 consecutive patients (training cohort, 197 patients; test cohort, 50 patients) with surgically proven gastric cancer. Dedicated radiomics prototype software was used to segment lesions on preoperative arterial phase (AP) CT images and extract features. A radiomics model was constructed to predict the LN metastasis by using a random forest (RF) algorithm. Finally, a nomogram was built incorporating the radiomics scores and selected clinical predictors. Receiver operating characteristic (ROC) curves were used to validate the capability of the radiomics model and nomogram on both the training and test cohorts. RESULTS: The radiomics model showed a favorable discriminatory ability in the training cohort with an area under the curve (AUC) of 0.844 (95% CI, 0.759 to 0.909), which was confirmed in the test cohort with an AUC of 0.837 (95% CI, 0.705 to 0.926). The nomogram consisted of radiomics scores and the CT-reported LN status showed excellent discrimination in the training and test cohorts with AUCs of 0.886 (95% CI, 0.808 to 0.941) and 0.881 (95% CI, 0.759 to 0.956), respectively. CONCLUSIONS: The CT-based radiomics nomogram holds promise for use as a noninvasive tool in the individual prediction of LN metastasis in gastric cancer. KEY POINTS: • CT radiomics showed a favorable performance for the prediction of LN metastasis in gastric cancer. • Radiomics model outperformed the routine CT in predicting LN metastasis in gastric cancer. • The radiomics nomogram holds potential in the individualized prediction of LN metastasis in gastric cancer.

Balance between Cytosolic and Chloroplast Translation Affects Leaf Variegation.

The development of functional chloroplasts relies on the fine coordination of expressions of both nuclear and chloroplast genomes. We have been using the Arabidopsis (Arabidopsis thaliana) yellow variegated (var2) leaf variegation mutant as a tool to dissect the regulation of chloroplast development. In this work, we screened for var2 genetic enhancer modifiers termed enhancer of variegation (evr) mutants and report the characterization of the first EVR locus, EVR1 We showed that EVR1 encodes the cytosolic 80S ribosome 40S small subunit protein RPS21B and the loss of EVR1 causes the enhancement of var2 leaf variegation. We further demonstrated that combined S21 activities from EVR1 and its close homolog, EVR1L1, are essential for Arabidopsis, and they act redundantly in regulating leaf development and var2 leaf variegation. Moreover, using additional cytosolic ribosomal protein mutants, we showed that although mutations in cytosolic ribosomal proteins all enhance var2 leaf variegation to varying degrees, the 40S subunit appears to have a more profound role over the 60S subunit in regulating VAR2-mediated chloroplast development. Comprehensive genetic analyses with var2 suppressors that are defective in chloroplast translation established that the enhancement of var2 leaf variegation by cytosolic ribosomal protein mutants is dependent on chloroplast translation. Based on our data, we propose a model that incorporates the suppression and enhancement of var2 leaf variegation, and hypothesize that VAR2/AtFtsH2 may be intimately involved in the balancing of cytosolic and chloroplast translation programs during chloroplast biogenesis.

Three-dimensional turbo-spin-echo amide proton transfer-weighted mri for cervical cancer: A preliminary study.

BACKGROUND: Amide proton transfer-weighted (APTw) imaging has shown great potential in the diagnosis of cancer, but has yet not been well studied in cervical cancer. PURPOSE: To evaluate the image quality and clinical feasibility of APTw MRI for cervical cancer. STUDY TYPE: Prospective. POPULATION: In all, 75 patients with cervical lesions and 49 healthy volunteers. FIELD STRENGTH/SEQUENCE: 3.0 T, 3D turbo spin echo (TSE) APTw sequence. ASSESSMENT: Three radiologists, blinded to the clinical data, independently evaluated APTw image quality with a 5-point Likert scale on 64 patients with pathologically confirmed cervical cancer. APT values, calculated based on asymmetry of acquired Z-spectrum with respect to water frequency, using 3D turbo spin echo volume acquisition with B0 correction, were independently measured by two radiologists, twice for each observer, on 52 cervical cancer lesions and 49 normal cervical stroma with a mean region of interest area of 638.6 mm2 and 557.5 mm2 , respectively. STATISTICAL TESTS: Interobserver agreement was evaluated by Kendall's W test. Intra- and interobserver interclass correlation coefficients (ICC) were computed. Student's t-test was used to compare the differences of APT values between cervical cancer and normal cervix; receiver operating characteristic analysis was performed. RESULTS: Most cases revealed good APTw image quality with excellent agreement (Kendall's W = 0.850, P < 0.001). APT values of cervical cancer and normal cervical stroma were 2.745 ± 0.065 and 1.853 ± 0.059, respectively, with a significant difference (P < 0.0001). Intraobserver ICCs were 0.963 and 0.960 for two readers. Interobserver ICC was 0.993. Area under the curve (AUC) for differentiating cervical cancer from normal cervical stroma was 0.927. The feasible threshold value for AUC was determined as 2.221 with sensitivity of 84.62% and specificity of 83.66%. DATA CONCLUSION: 3D TSE APTw MRI is feasible in cervical cancer. Cervical cancer showed significantly higher APT values than normal cervix. LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;50:1318-1325.

A Putative Chloroplast Thylakoid Metalloprotease VIRESCENT3 Regulates Chloroplast Development in Arabidopsis thaliana.

The chloroplast is the site of photosynthesis and many other essential plant metabolic processes, and chloroplast development is an integral part of plant growth and development. Mutants defective in chloroplast development can display various color phenotypes including the intriguing virescence phenotype, which shows yellow/white coloration at the leaf base and greening toward the leaf tip. Through large scale genetic screens, we identified a series of new virescent mutants including virescent3-1 (vir3-1), vir4-1, and vir5-1 in Arabidopsis thaliana. We showed that VIR3 encodes a putative chloroplast metalloprotease by map-based cloning. Through site-directed mutagenesis, we showed that the conserved histidine 235 residue in the zinc binding motif HEAGH of VIR3 is indispensable for VIR3 accumulation in the chloroplast. The chloroplast localization of VIR3 was confirmed by the transient expression of VIR3-GFP in leaf protoplasts. Furthermore, taking advantage of transgenic lines expressing VIR3-FLAG, we demonstrated that VIR3 is an intrinsic thylakoid membrane protein that mainly resides in the stromal lamellae. Moreover, topology analysis using transgenic lines expressing a dual epitope-tagged VIR3 indicated that both the N and C termini of VIR3 are located in the stroma, and the catalytic domain of VIR3 is probably facing the stroma. Blue native gel analysis indicated that VIR3 is likely present as a monomer or part of a small complex in the thylakoid membrane. This work not only implicates VIR3 as a new factor involved in early chloroplast development but also provides more insight into the roles of chloroplast proteases in chloroplast biogenesis.