Global hyperperfusion after successful endovascular thrombectomy is linked to worse outcome in acute ischemic stroke.

Patients with stroke may develop hyperperfusion after a successful endovascular thrombectomy (EVT). However, the relationship between post-EVT hyperperfusion and clinical outcomes remains unclear and requires further clarification. We reviewed consecutive patients with anterior circulation occlusion who were successfully recanalized with EVT. Based on post-EVT arterial spin-labeling images, hyperperfusion was categorized as follows: global hyperperfusion (GHP), increased cerebral blood flow (CBF) in ≥ 50% of the culprit vessel territory; focal hyperperfusion (FHP), increased CBF in < 50% of the culprit vessel territory; no hyperperfusion (NHP), no discernible CBF increase. Factors associated with hyperperfusion were assessed, and clinical outcomes were compared among patients under different hyperperfusion categories. Among 131 patients, 25 and 40 patients developed GHP and FHP, respectively. Compared to other groups, the GHP group had worse National Institutes of Health Stroke Scale score (GHP vs. NHP/FHP, 18.1 ± 7.4 vs. 12.3 ± 6.0; p < 0.001), a larger post-EVT infarct volume (98.9 [42.3-132.7] vs. 13.5 [5.0-34.1] mL; p < 0.001), and a worse 90-day outcome (modified Rankin Scale, 3 [1-4] vs. 2 [0-3]; p = 0.030). GHP was independently associated with infarct volume (B = 0.532, standard error = 0.163, p = 0.001), and infarct volume was a major mediator of the association of GHP with unfavorable outcomes (total effect: ß = 0.176, p = 0.034; direct effect: ß = 0.045, p = 0.64; indirect effect: ß = 0.132, p = 0.017). Patients presenting with post-EVT GHP had poorer neurological prognosis, which is likely mediated by a large infarct volume.

Scientific Publications on Thyroid Ultrasound between 2001 and 2020: Differences in Research Characteristics by Disciplines.

OBJECTIVE: To analyze the characteristics and trends of scientific publications on thyroid ultrasound (US) from 2001 to 2020, specifically examining the differences among disciplines. MATERIALS AND METHODS: The MEDLINE database was searched for scientific articles on thyroid US published between 2001 and 2020 using the PubMed online service. The evaluated parameters included year of publication, type of document, topic, funding, first author's specialty, journal name, subject category, impact factor, and quartile ranking of the publishing journal, country, and language. Relationships between the first author's specialty (radiology, internal medicine, surgery, otorhinolaryngology, and miscellaneous) and other parameters were analyzed. RESULTS: A total of 2917 thyroid US publications were published between 2001 and 2020, which followed an exponential growth pattern, with an annual growth rate of 11.6%. Radiology produced the most publications (n = 1290, 44.2%), followed by internal medicine (n = 716, 24.5%), surgery (n = 409, 14.0%), and otorhinolaryngology (n = 171, 5.9%). Otorhinolaryngology and internal medicine published significantly more case reports than radiology (p < 0.001, each). Radiology published a significantly higher proportion of publications on imaging diagnosis (p < 0.001 for all) and a significantly lower proportion of publications on biopsy (p < 0.001 for all) than the other disciplines. Publications produced by radiology authors were less frequently published in Q1 journals than those from other disciplines (p < 0.005 for internal medicine and miscellaneous disciplines and < 0.01 for surgery and otorhinolaryngology). China contributed the greatest number of publications (n = 622, 21.3%), followed by South Korea (n = 478, 16.4%) and the United States (n = 468, 16.0%). CONCLUSION: Radiology produced the most publications for thyroid US than any other discipline. Radiology authors published more notably on imaging diagnosis compared to other topics and in journals with lower impact factors compared to authors in other disciplines.

SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence.

Alternative splicing (AS) is a procedure during gene expression that allows the production of multiple mRNAs from a single gene, leading to a larger number of proteins with various functions. The alternative splicing (AS) of Fas (Apo-1/CD95) pre-mRNA can generate membrane-bound or soluble isoforms with pro-apoptotic and anti-apoptotic functions. SRSF6, a member of the Serine/Arginine-rich protein family, plays essential roles in both constitutive and alternative splicing. Here, we identified SRSF6 as an important regulatory protein in Fas AS. The cassette exon inclusion of Fas was decreased by SRSF6-targeting shRNA treatment, but increased by SRSF6 overexpression. The deletion and substitution mutagenesis of the Fas minigene demonstrated that the UGCCAA sequence in the cassette exon of the Fas gene causes the functional disruption of SRSF6, indicating that these sequences are essential for SRSF6 function in Fas splicing. In addition, biotin-labeled RNA-pulldown and immunoblotting analysis showed that SRSF6 interacted with these RNA sequences. Mutagenesis in the splice-site strength alteration demonstrated that the 5' splice-site, but not the 3' splice-site, was required for the SRSF6 regulation of Fas pre-mRNA. In addition, a large-scale RNA-seq analysis using GTEX and TCGA indicated that while SRSF6 expression was correlated with Fas expression in normal tissues, the correlation was disrupted in tumors. Furthermore, high SRSF6 expression was linked to the high expression of pro-apoptotic and immune activation genes. Therefore, we identified a novel RNA target with 5' splice-site dependence of SRSF6 in Fas pre-mRNA splicing, and a correlation between SRSF6 and Fas expression.

Comparison of gadoxetate disodium-enhanced MRI sequences for measuring hepatic observation size and its implication of LI-RADS classification.

PURPOSE: We aimed to determine the optimal image sequence for measurement of hepatic observations on gadoxetate disodium-enhanced MRI in comparison with pathologic measurement, and to evaluate its clinical impact on the Liver Imaging Reporting and Data System (LI-RADS) v2018 classification. METHODS: Two hundred and fifty-three patients (279 hepatic observations) who underwent gadoxetate disodium-enhanced MRI and subsequent hepatectomy were retrospectively included. Two radiologists independently evaluated the visualization score (five-point scale) and size of each observation on six MRI sequences (T1-weighted, T2-weighted, arterial-phase, portal venous-phase, transitional-phase [TP], and hepatobiliary-phase [HBP] images) and assigned a LI-RADS category. Correlations between MRI and pathologic measurements were evaluated using Pearson correlation coefficients. A repeated measures analysis of variance with Bonferroni post hoc comparison tests was used to compare the visualization scores and absolute differences between MRI sequences and pathologic measurements. The LI-RADS classification according the size measurement of each MRI sequence was compared using Cochran's Q test with a post hoc McNemar's test. RESULTS: Of the MRI sequences, HBP had the highest visualization score (4.1 ± 0.6) and correlation coefficient (r = 0.965). The absolute difference between MRI and pathologic measurement was lowest on TP (2.3 mm ± 2.2), followed by HBP (2.4 mm ± 2.1). In the LI-RADS classifications, HBP did not have any non-visible observations. Regarding LR-3, LR-4, and LR-5, there was no significantly different LI-RADS classification among the six MRI sequences (p ≥ 0.122). CONCLUSION: Hepatobiliary-phase images are clinically useful for measuring hepatic observations on gadoxetate disodium-enhanced MRI, especially regarding visibility and correlation with pathologic findings.

MRI features for differentiation of autoimmune pancreatitis from pancreatic ductal adenocarcinoma: A systematic review and meta-analysis.

BACKGROUND AND AIMS: The accurate differential diagnosis between autoimmune pancreatitis (AIP) and pancreatic ductal adenocarcinoma (PDAC) is clinically important. We aimed to determine significant MRI features for differentiating AIP from PDAC, including assessment of diffusion-weighted imaging (DWI). METHODS: We performed a systematic search using three databases. The pooled diagnostic odds ratio was calculated using a bivariate random effects model to determine significant MRI features for differentiating AIP from PDAC. The pooled sensitivity and specificity were calculated. The qualitative systematic review for DWI assessment was performed. RESULTS: Of nine studies (775 patients), multiple main pancreatic duct (MPD) strictures, absence of upstream marked MPD dilatation, peripancreatic rim, and duct penetration sign were significant MRI features for differentiating AIP from PDAC. Absence of MPD dilatation had the highest pooled sensitivity (87%, 95% CI=68-96%), whereas peripancreatic rim had the highest pooled specificity (100%, 95% CI=88-100%). Of 12 studies evaluating DWI, seven reported statistically significant differences in apparent diffusion coefficient (ADC) values between AIP and PDAC; however, four reported lower ADC values in AIP than in PDAC, but three reported the opposite result. CONCLUSION: The four significant MRI features can be useful to differentiate AIP from PDAC, but DWI assessment might be limited.

Is the Mixed Use of Magnetic Resonance Enterography and Computed Tomography Enterography Adequate for Routine Periodic Follow-Up of Bowel Inflammation in Patients with Crohn's Disease?

OBJECTIVE: Computed tomography enterography (CTE) and magnetic resonance enterography (MRE) are considered substitutes for each other for evaluating Crohn's disease (CD). However, the adequacy of mixing them for routine periodic follow-up for CD has not been established. This study aimed to compare MRE alone with the mixed use of CTE and MRE for the periodic follow-up of small bowel inflammation in patients with CD. MATERIALS AND METHODS: We retrospectively compared two non-randomized groups, each comprising 96 patients with CD. One group underwent CTE and MRE (MRE followed by CTE or vice versa) for the follow-up of CD (interval, 13-27 months [median, 22 months]), and the other group underwent MRE alone (interval, 15-26 months [median, 21 months]). However, these two groups were similar in clinical characteristics. Three independent readers from three different institutions determined whether inflammation had decreased, remained unchanged, or increased within the entire small bowel and the terminal ileum based on sequential enterography of the patients after appropriate blinding. We compared the two groups for inter-reader agreement and accuracy (terminal ileum only) using endoscopy as the reference standard for enterographic interpretation. RESULTS: The inter-reader agreement was greater in the MRE alone group for the entire small bowel (intraclass correlation coefficient [ICC]: 0.683 vs. 0.473; p = 0.005) and the terminal ileum (ICC: 0.656 vs. 0.490; p = 0.030). The interpretation accuracy was higher in the MRE alone group without statistical significance (70.9%-74.5% vs. 57.9%-64.9% in individual readers; adjusted odds ratio = 3.21; p = 0.077). CONCLUSION: The mixed use of CTE and MRE was inferior to MRE alone in terms of inter-reader reliability and could probably be less accurate than MRE alone for routine monitoring of small bowel inflammation in patients with CD. Therefore, the consistent use of MRE is favored for this purpose.

Global Alternative Splicing Defects in Human Breast Cancer Cells.

Breast cancer is the most frequently occurred cancer type and the second cause of death in women worldwide. Alternative splicing (AS) is the process that generates more than one mRNA isoform from a single gene, and it plays a major role in expanding the human protein diversity. Aberrant AS contributes to breast cancer metastasis and resistance to chemotherapeutic interventions. Therefore, identifying cancer-specific isoforms is the prerequisite for therapeutic interventions intended to correct aberrantly expressed AS events. Here, we performed RNA-mediated oligonucleotide annealing, selection, and ligation coupled with next-generation sequencing (RASL-seq) in breast cancer cells, to identify global breast cancer-specific AS defects. By RT-PCR validation, we demonstrate the high accuracy of RASL-seq results. In addition, we analyzed identified AS events using the Cancer Genome Atlas (TCGA) database in a large number of non-pathological and breast tumor specimens and validated them in normal and breast cancer samples. Interestingly, aberrantly regulated AS cassette exons in cancer tissues do not encode for known functional domains but instead encode for amino acids constituting regions of intrinsically disordered protein portions characterized by high flexibility and prone to be subjected to post-translational modifications. Collectively, our results reveal novel AS errors occurring in human breast cancer, potentially affecting breast cancer-related biological processes.

Widespread Alternative Splicing Changes in Metastatic Breast Cancer Cells.

Aberrant alternative splicing (AS) is a hallmark of cancer and a potential target for novel anti-cancer therapeutics. Breast cancer-associated AS events are known to be linked to disease progression, metastasis, and survival of breast cancer patients. To identify altered AS programs occurring in metastatic breast cancer, we perform a global analysis of AS events by using RNA-mediated oligonucleotide annealing, selection, and ligation coupled with next-generation sequencing (RASL-seq). We demonstrate that, relative to low-metastatic, high-metastatic breast cancer cells show different AS choices in genes related to cancer progression. Supporting a global reshape of cancer-related splicing profiles in metastatic breast cancer we found an enrichment of RNA-binding motifs recognized by several splicing regulators, which have aberrant expression levels or activity during breast cancer progression, including SRSF1. Among SRSF1-regulated targets we found DCUN1D5, a gene for which skipping of exon 4 in its pre-mRNA introduces a premature termination codon (PTC), thus generating an unstable transcript degraded by nonsense-mediated mRNA decay (NMD). Significantly, distinct breast cancer subtypes show different DCUN1D5 isoform ratios with metastatic breast cancer expressing the highest level of the NMD-insensitive DCUN1D5 mRNA, thus showing high DCUN1D5 expression levels, which are ultimately associated with poor overall and relapse-free survival in breast cancer patients. Collectively, our results reveal global AS features of metastatic breast tumors, which open new possibilities for the treatment of these aggressive tumor types.

SRSF9 Regulates Cassette Exon Splicing of Caspase-2 by Interacting with Its Downstream Exon.

Alternative splicing (AS) is an important posttranscriptional regulatory process. Damaged or unnecessary cells need to be removed though apoptosis to maintain physiological processes. Caspase-2 pre-mRNA produces pro-apoptotic long mRNA and anti-apoptotic short mRNA isoforms through AS. How AS of Caspase-2 is regulated remains unclear. In the present study, we identified a novel regulatory protein SRSF9 for AS of Caspase-2 cassette exon 9. Knock-down (KD) of SRSF9 increased inclusion of cassette exon and on the other hand, overexpression of SRSF9 decreased inclusion of this exon. Deletion mutagenesis demonstrated that exon 9, parts of intron 9, exon 8 and exon 10 were not required for the role of SRSF9 in Caspase-2 AS. However, deletion and substitution mutation analysis revealed that AGGAG sequence located at exon 10 provided functional target for SRSF9. In addition, RNA-pulldown mediated immunoblotting analysis showed that SRSF9 interacted with this sequence. Gene ontology analysis of RNA-seq from SRSF9 KD cells demonstrates that SRSF9 could regulate AS of a subset of apoptosis related genes. Collectively, our results reveal a basis for regulation of Caspase-2 AS.

Meta-analysis of CT and MRI for differentiation of autoimmune pancreatitis from pancreatic adenocarcinoma.

OBJECTIVES: To systematically determine the diagnostic performance of computed tomography (CT) and magnetic resonance imaging (MRI) for differentiating autoimmune pancreatitis (AIP) from pancreatic ductal adenocarcinoma (PDAC), with a comparison between the two imaging modalities. METHODS: Literature search was conducted using PubMed and EMBASE databases to identify original articles published between 2009 and 2019 reporting the diagnostic performance of CT and MRI for differentiating AIP from PDAC. The meta-analytic sensitivity and specificity of CT and MRI were calculated, and compared using a bivariate random effects model. Subgroup analysis for differentiating focal AIP from PDAC was performed. RESULTS: Of the 856 articles screened, 11 eligible articles are remained, i.e., five studies for CT, four for MRI, and two for both. The meta-analytic summary sensitivity and specificity of CT were 59% (95% confidence interval [CI], 41-75%) and 99% (95% CI, 88-100%), respectively, while those of MRI were 84% (95% CI, 68-93%) and 97% (95% CI, 87-99%). MRI had a significantly higher meta-analytic summary sensitivity than CT (84% vs. 59%, p = 0.02) but a similar specificity (97% vs. 99%, p = 0.18). In the subgroup analysis for focal AIP, the sensitivity for distinguishing between focal AIP and PDAC was lower than that for the overall analysis. MRI had a higher sensitivity than CT (76% vs. 50%, p = 0.28) but a similar specificity (97% vs. 98%, p = 0.07). CONCLUSION: MRI might be clinically more useful to evaluate patients with AIP, particularly for differentiating AIP from PDAC. KEY POINTS: • MRI had an overall good diagnostic performance to differentiate AIP from PDAC with a meta-analytic summary estimate of 83% for sensitivity and of 97% for specificity. • CT had a very high specificity (99%), but a suboptimal sensitivity (59%) for differentiating AIP from PDAC. • Compared with CT, MRI had a higher sensitivity, but a similar specificity.

U2AF65-Dependent SF3B1 Function in SMN Alternative Splicing.

Splicing factor 3b subunit 1 (SF3B1) is an essential protein in spliceosomes and mutated frequently in many cancers. While roles of SF3B1 in single intron splicing and roles of its cancer-linked mutant in aberrant splicing have been identified to some extent, regulatory functions of wild-type SF3B1 in alternative splicing (AS) are not well-understood yet. Here, we applied RNA sequencing (RNA-seq) to analyze genome-wide AS in SF3B1 knockdown (KD) cells and to identify a large number of skipped exons (SEs), with a considerable number of alternative 5' splice-site selection, alternative 3' splice-site selection, mutually exclusive exons (MXE), and retention of introns (RI). Among altered SEs by SF3B1 KD, survival motor neuron 2 (SMN2) pre-mRNA exon 7 splicing was a regulatory target of SF3B1. RT-PCR analysis of SMN exon 7 splicing in SF3B1 KD or overexpressed HCT116, SH-SY5Y, HEK293T, and spinal muscular atrophy (SMA) patient cells validated the results. A deletion mutation demonstrated that the U2 snRNP auxiliary factor 65 kDa (U2AF65) interaction domain of SF3B1 was required for its function in SMN exon 7 splicing. In addition, mutations to lower the score of the polypyrimidine tract (PPT) of exon 7, resulting in lower affinity for U2AF65, were not able to support SF3B1 function, suggesting the importance of U2AF65 in SF3B1 function. Furthermore, the PPT of exon 7 with higher affinity to U2AF65 than exon 8 showed significantly stronger interactions with SF3B1. Collectively, our results revealed SF3B1 function in SMN alternative splicing.

Opposite Roles of Tra2ß and SRSF9 in the v10 Exon Splicing of CD44.

CD44 is a transmembrane glycoprotein involved in cell-cell and cell-matrix interactions. Several CD44 protein isoforms are generated in human through alternative splicing regulation of nine variable exons encoding for the extracellular juxta-membrane region. While the CD44 splicing variants have been described to be involved in cancer progression and development, the regulatory mechanism(s) underlying their production remain unclear. Here, we identify Tra2ß and SRSF9 as proteins with opposite roles in regulating CD44 exon v10 splicing. While Tra2ß promotes v10 inclusion, SRSF9 inhibits its inclusion. Mechanistically, we found that both proteins are able to target v10 exon, with GAAGAAG sequence being the binding site for Tra2ß and AAGAC that for SRSF9. Collectively, our data add a novel layer of complexity to the sequential series of events involved in the regulation of CD44 splicing.

RRM but not the Asp/Glu domain of hnRNP C1/C2 is required for splicing regulation of Ron exon 11 pre-mRNA.

The Ron proto-oncogene is a human receptor for macrophage-stimulating protein (MSP). The exclusion of exon 11 in alternative splicing generates ΔRON protein that is constitutively activated. Heterogenous ribonucleaoprotein (hnRNP) C1/C2 is one of the most abundant proteins in cells. In this manuscript, we showed that both hnRNP C1 and C2 promoted exon 11 inclusion of Ron pre-mRNA and that hnRNP C1 and hnRNP C2 functioned independently but not cooperatively. Moreover, hnRNP C1 stimulated exon 11 splicing through intron 10 activation but not through intron 11 splicing. Furthermore, we showed that, whereas the RRM domain was required for hnRNP C1 function, the Asp/Glu domain was not. In conclusion, hnRNP C1/C2 promoted exon 11 splicing independently by stimulating intron 10 splicing through RRM but not through the Asp/Glu domain. [BMB Reports 2019; 52(11): 641-646].

Activation of Cryptic 3' Splice-Sites by SRSF2 Contributes to Cassette Exon Skipping.

Here we show that the serine/arginine rich splicing factor 2 (SRSF2) promotes cryptic 3' splice-site (3'AG') usage during cassette exon exclusion in survival of motor neuron (SMN2) minigenes. Deletion of the 3'AG' (3'AG'1), its associated branch point (BP') and polypyrimidine tract (PPT') sequences directs SRSF2 to promote a second 3'AG' (3'AG'2) with less conserved associated region for intron splicing. Furthermore, deletion of both 3'AG'1 and 3'AG'2 and their associated sequences triggered usage of a third 3'AG'3 that has very weak associated sequences. Interestingly, when intron splicing was directed to the 3'AG' cryptic splice-sites, intron splicing from the canonical 3'AG splice-site was reduced along with a decrease in cassette exon inclusion. Moreover, multiple SRSF2 binding sites within the intron are responsible for 3'AG' activation. We conclude that SRSF2 facilitates exon exclusion by activating a cryptic 3'AG' and inhibiting downstream intron splicing.

Binding of SRSF4 to a novel enhancer modulates splicing of exon 6 of Fas pre-mRNA.

Alternative splicing of exon 6 in Fas pre-mRNA generates a membrane bound pro-apoptotic isoform or soluble anti-apoptotic isoform. SRSF4 is a member of Arginine-Serine rich (SR) protein family. Here we demonstrate that increased SRSF4 expression stimulates exon 6 inclusion, and that reduced SRSF4 expression promotes exon 6 exclusion. We also show that weaker but not stronger 5' splice-site strength of exon 6 abolishes the SRSF4 effects on exon 6 splicing. Furthermore, we identified a novel enhancer on exon 6, on which SRSF4 interacts functionally and physically. Our results illustrate a novel regulatory mechanism of Fas pre-mRNA splicing.