Το μέτρο Brown για μη κανονικούς τελεστές και η οριακή θεωρία τυχαίων πινάκων

Η εργασία ασχολείται με τη μελέτη του μέτρου Brown ορισμένων μη ερμιτιανών τελεστών που προκύπτουν από τη θεωρία ελεύθερης πιθανότητας του Voiculescu. Συνήθως αυτοί οι τελεστές εμφανίζονται ως το όριο των ⋆-ροπών ορισμένων συνόλων μη ερμιτιανών τυχαίων πινάκων και το Brown μέτρο δίνει τότε έναν κανονικό υποψήφιο για την οριακή κατανομή ιδιοτιμών των τυχαίων πινάκων. Μια σημαντική κλάση τελεστών που θα μας απασχολήσει είναι τα πολυώνυμα σε ⋆-ελεύθερες μεταβλητές. Άλλα παραδείγματα αποτελούν τα R-διαγώνια στοιχεία και τα ελλειπτικά στοιχεία, για τα οποία ήταν ήδη γνωστό το μέτρο Brown, καθώς και μια νέα κατηγορία τριγωνικών - ελλειπτικών στοιχείων. Η μέθοδός μας για τον υπολογισμό του μέτρου Brown συνδυάζει τη μέθοδο της ερμιτιανής αναγωγής, όπως εξετάζεται στη φυσική βιβλιογραφία, με το τέχνασμα της γραμμικοποίησης και της μείωσης της τάξης, και επιτρέπει λεπτομερείς υπολογισμούς.We study the Brown measure of certain non–Hermitian operators arising from Voiculescu’s free probability theory. Usually those operators appear as the limit in ⋆-moments of certain ensembles of non–Hermitian random matrices, and the Brown measure gives then a canonical candidate for the limit eigenvalue distribution of the random matrices. A prominent class for our operators is given by polynomials in ⋆-free variables. Other explicit examples include R–diagonal elements and elliptic elements, for which the Brown measure was already known, and a new class of triangular–elliptic elements. Our method for the calculation of the Brown measure is based on a rigorous mathematical treatment of the Hermitian reduction method, as considered in the physical literature, combined with subordination and the linearization trick

Zebrafish models for ARVC8 analysis and drug discovery

INTRODUCTION: Desmoplakin is one the most abundant desmosomal proteins in cardiac and epithelial tissues. In humans, dominat mutations in the desmoplakin gene (DSP) cause Arrhythmogenic Right Ventricular Cardio​myopathy 8 (ARVC8), a dominant cardiomyopathy, frequently involved in juvenile sudden death. Current ARVC models are based on cell lines and transgenic mice. In this context, it has been shown that suppression of DSP expression leads to a reduction in canonical Wnt signaling, suggesting that this pathway could be a molecular target for ARVC therapeutic intervention. In order to address this issue, the present study aims to evaluate the pathogenic mechanisms of DSP mutations in vivo, using zebrafish (Danio rerio) as an innovative model for this disease. In zebrafish, the desmoplakin gene is present with two isoforms, dspa and dspb, both orthologous to the single DSP in humans. PURPOSE: The purpose of this study is the generation and the phenotypic characterization of transient ARVC8 zebrafish models using a morpholino-mediated knock-down strategy. In addition, by taking advantage of zebrafish pathway reporter lines, we aim to verify if Wnt signaling and/or other molecular cascades might be involved in ARVC8 pathogenesis. The final goal is the assessment of our ARVC8 model as a suitable tool for molecularly-targeted drug discovery. METHODS: To evaluate the expression of dspa and dspb during zebrafish embryonic development and adulthood, we used whole-mount in situ hybridization (WISH) and semi quantitative RT_PCR. Knockdown of zebrafish dspa and dspb genes was obtained by a morpholino (MO)-based antisense strategy. Specifically, we injected anti-dspa and anti-dspb MO oligos in both wild types and pathway-specific lines reporting the activity of Wnt, Bmp, TGFbeta, FGF, Shh, Notch, CREB, Hippo and Hypoxia signaling. RESULTS: We found that both dspa and dspb are expressed during zebrafish embryonic development, while the molecular analysis of cDNAs from different adult tissues demonstrates that both dspa and dspb are highly expressed in heart and skin, with dspa more strongly detectable compared to dspb. MO-mediated knock-down of both dspa and dspb leads to delayed development, microcephaly, pericardial edema and, particularly in dspb knock-down embryos, decreased heart rate. TEM analysis of cardiac and skin tissues under dspa+dspb simultaneous knock-down shows reduced and disorganized desmososmes. As far as concerns the analysis of previously mentioned signaling pathways, we observed a specific reduction of Wnt signaling responsiveness in the cardiac region of both dspa and dspb knock- down embryos (Fig. 1). CONCLUSION: Our results show that transient knock-down of zebrafish desmoplakin genes is able to phenocopy some ARVC8 features, such as cardiac and cutaneous desmosomal defects, heart rate alteration and Wnt signaling reduction, pointing to zebrafish as a suitable ARVC8 model for in vivo screening of molecularly-targeted drugs

A microRNA Expression Profile as Non-Invasive Biomarker in a Large Arrhythmogenic Cardiomyopathy Cohort

Arrhythmogenic Cardiomyopathy (AC) is a clinically and genetically heterogeneous myocardial disease. Half of AC patients harbour private desmosomal gene variants. Although microRNAs (miRNAs) have emerged as key regulator molecules in cardiovascular diseases and their involvement, correlated to phenotypic variability or to non-invasive biomarkers, has been advanced also in AC, no data are available in larger disease cohorts. Here, we propose the largest AC cohort unbiased by technical and biological factors. MiRNA profiling on nine right ventricular tissue, nine blood samples of AC patients, and four controls highlighted 10 differentially expressed miRNAs in common. Six of these were validated in a 90-AC patient cohort independent from genetic status: miR-122-5p, miR-133a-3p, miR-133b, miR-142-3p, miR-182-5p, and miR-183-5p. This six-miRNA set showed high discriminatory diagnostic power in AC patients when compared to controls (AUC-0.995), non-affected family members of AC probands carrying a desmosomal pathogenic variant (AUC-0.825), and other cardiomyopathy groups (Hypertrophic Cardiomyopathy: AUC-0.804, Dilated Cardiomyopathy: AUC-0.917, Brugada Syndrome: AUC-0.981, myocarditis: AUC-0.978). AC-related signalling pathways were targeted by this set of miRNAs. A unique set of six-miRNAs was found both in heart-tissue and blood samples of AC probands, supporting its involvement in disease pathogenesis and its possible role as a non-invasive AC diagnostic biomarker

Arrhythmogenic Right Ventricular Cardiomyopathy: Characterization of Left Ventricular Phenotype and Differential Diagnosis With Dilated Cardiomyopathy

Background This study assessed the prevalence of left ventricular (LV) involvement and characterized the clinical, electrocardiographic, and imaging features of LV phenotype in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC). Differential diagnosis between ARVC-LV phenotype and dilated cardiomyopathy (DCM) was evaluated. Methods and Results The study population included 87 ARVC patients (median age 34\ua0years) and 153 DCM patients (median age 51\ua0years). All underwent cardiac magnetic resonance with quantitative tissue characterization. Fifty-eight ARVC patients (67%) had LV involvement, with both LV systolic dysfunction and LV late gadolinium enhancement (LGE) in 41/58 (71%) and LV-LGE in isolation in 17 (29%). Compared with DCM, the ARVC-LV phenotype was statistically significantly more often characterized by low QRS voltages in limb leads, T-wave inversion in the inferolateral leads and major ventricular arrhythmias. LV-LGE was found in all ARVC patients with LV systolic dysfunction and in 69/153 (45%) of DCM patients. Patients with ARVC and LV systolic dysfunction had a greater amount of LV-LGE (25% versus 13% of LV mass; P<0.01), mostly localized in the subepicardial LV wall layers. An LV-LGE 6520% had a 100% specificity for diagnosis of ARVC-LV phenotype. An inverse correlation between LV ejection fraction and LV-LGE extent was found in the ARVC-LV phenotype (r=-0.63; P<0.01), but not in DCM (r=-0.01; P=0.94). Conclusions LV involvement in ARVC is common and characterized by clinical and cardiac magnetic resonance features which differ from those seen in DCM. The most distinctive feature of ARVC-LV phenotype is the large amount of LV-LGE/fibrosis, which impacts directly and negatively on the LV systolic function

Morphofunctional abnormalities of mitral annulus and arrhythmic mitral valve prolapse

Background\u2014Arrhythmic mitral valve prolapse (MVP) is characterized by myxomatous leaflets and left ventricular (LV) fibrosis of papillary muscles and inferobasal wall. We searched for morphofunctional abnormalities of the mitral valve that could explain a regional mechanical myocardial stretch. Methods and Results\u2014Thirty-six (27 female patients; median age: 44 years) arrhythmic MVP patients with LV late gadolinium enhancement on cardiac magnetic resonance and no or trivial mitral regurgitation, and 16 (6 female patients; median age: 40 years) MVP patients without LV late gadolinium enhancement were investigated by morphofunctional cardiac magnetic resonance. Mitral annulus disjunction (median: 4.8 versus 1.8 mm; P1.5 (22 [61%] versus 4 [25%]; P=0.016) were higher in MVP patients with late gadolinium enhancement than in those without. A linear correlation was found between mitral annulus disjunction and curling (R=0.85). A higher prevalence of auscultatory midsystolic click (26 [72%] versus 6 [38%]; P=0.018) was also noted. Histology of the mitral annulus showed a longer mitral annulus disjunction in 50 sudden death patients with MVP and LV fibrosis than in 20 patients without MVP (median: 3 versus 1.5 mm; P<0.001). Conclusions\u2014Mitral annulus disjunction is a constant feature of arrhythmic MVP with LV fibrosis. The excessive mobility of the leaflets caused by posterior systolic curling accounts for a mechanical stretch of the inferobasal wall and papillary muscles, eventually leading to myocardial hypertrophy and scarring. These mitral annulus abnormalities, together with auscultatory midsystolic click, may identify MVP patients who would need arrhythmic risk stratification

A novel DSP zebrafish model reveals training- and drug-induced modulation of arrhythmogenic cardiomyopathy phenotypes

Arrhythmogenic cardiomyopathy (AC) is an inherited disorder characterized by progressive loss of the ventricular myocardium causing life-threatening ventricular arrhythmias, syncope and sudden cardiac death in young and athletes. About 40% of AC cases carry one or more mutations in genes encoding for desmosomal proteins, including Desmoplakin (Dsp). We present here the first stable Dsp knock-out (KO) zebrafish line able to model cardiac alterations and cell signalling dysregulation, characteristic of the AC disease, on which environmental factors and candidate drugs can be tested. Our stable Dsp knock-out (KO) zebrafish line was characterized by cardiac alterations, oedema and bradycardia at larval stages. Histological analysis of mutated adult hearts showed reduced contractile structures and abnormal shape of the ventricle, with thinning of the myocardial layer, vessels dilation and presence of adipocytes within the myocardium. Moreover, TEM analysis revealed “pale”, disorganized and delocalized desmosomes. Intensive physical training protocol caused a global worsening of the cardiac phenotype, accelerating the progression of the disease. Of note, we detected a decrease of Wnt/β-catenin signalling, recently associated with AC pathogenesis, as well as Hippo/YAP-TAZ and TGF-β pathway dysregulation. Pharmacological treatment of mutated larvae with SB216763, a Wnt/β-catenin agonist, rescued pathway expression and cardiac abnormalities, stabilizing the heart rhythm. Overall, our Dsp KO zebrafish line recapitulates many AC features observed in human patients, pointing at zebrafish as a suitable system for in vivo analysis of environmental modulators, such as the physical exercise, and the screening of pathway-targeted drugs, especially related to the Wnt/β-catenin signalling cascade

Circulating Cell-Free DNA in Dogs with Mammary Tumors: Short and Long Fragments and Integrity Index

Circulating cell-free DNA (cfDNA) has been considered an interesting diagnostic/prognostic plasma biomarker in tumor-bearing subjects. In cancer patients, cfDNA can hypothetically derive from tumor necrosis/apoptosis, lysed circulating cells, and some yet unrevealed mechanisms of active release. This study aimed to preliminarily analyze cfDNA in dogs with canine mammary tumors (CMTs). Forty-four neoplastic, 17 non-neoplastic disease-bearing, and 15 healthy dogs were recruited. Necrosis and apoptosis were also assessed as potential source of cfDNA on 78 CMTs diagnosed from the 44 dogs. The cfDNA fragments and integrity index significantly differentiated neoplastic versus non-neoplastic dogs (P<0.05), and allowed the distinction between benign and malignant lesions (P<0.05). Even if without statistical significance, the amount of cfDNA was also affected by tumor necrosis and correlated with tumor size and apoptotic markers expression. A significant (P<0.01) increase of Bcl-2 in malignant tumors was observed, and in metastatic CMTs the evasion of apoptosis was also suggested. This study, therefore, provides evidence that cfDNA could be a diagnostic marker in dogs carrying mammary nodules suggesting that its potential application in early diagnostic procedures should be further investigated

International Evidence Based Reappraisal of Genes Associated With Arrhythmogenic Right Ventricular Cardiomyopathy Using the Clinical Genome Resource Framework

Background - Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited disease characterized by ventricular arrhythmias and progressive ventricular dysfunction. Genetic testing is recommended and a pathogenic variant in an ARVC-associated gene is a major criterion for diagnosis according to the 2010 Task Force Criteria (TFC). As incorrect attribution of a gene to ARVC can contribute to misdiagnosis, we assembled an international multidisciplinary ARVC ClinGen Gene Curation Expert Panel to reappraise all reported ARVC genes. / Methods - Following a comprehensive literature search, six two-member teams conducted blinded independent curation of reported ARVC genes using the semi-quantitative ClinGen framework. /Results - Of 26 reported ARVC genes, only six (PKP2, DSP, DSG2, DSC2, JUP, TMEM43) had strong evidence and were classified as definitive for ARVC causation. There was moderate evidence for two genes, DES and PLN. The remaining 18 genes had limited or no evidence. RYR2 was refuted as an ARVC gene since clinical data and model systems exhibited a catecholaminergic polymorphic ventricular tachycardia (CPVT) phenotype. In ClinVar, only 5 pathogenic / likely pathogenic (P/LP) variants (1.1%) in limited evidence genes had been reported in ARVC cases in contrast to 450 desmosome gene variants (97.4%). / Conclusions - Using the ClinGen approach to gene-disease curation, only eight genes, (PKP2, DSP, DSG2, DSC2, JUP, TMEM43, PLN, DES) had definitive or moderate evidence for ARVC and these genes accounted for nearly all P/LP ARVC variants in ClinVar. Therefore, only P/LP variants in these eight genes should yield a major criterion for ARVC diagnosis. P/LP variants identified in other genes in a patient should prompt further phenotyping as variants in many of these genes are associated with other cardiovascular conditions