Utility of Post-Mortem Genetic Testing in Cases of Sudden Arrhythmic Death Syndrome.

BACKGROUND: Sudden arrhythmic death syndrome (SADS) describes a sudden death with negative autopsy and toxicological analysis. Cardiac genetic disease is a likely etiology. OBJECTIVES: This study investigated the clinical utility and combined yield of post-mortem genetic testing (molecular autopsy) in cases of SADS and comprehensive clinical evaluation of surviving relatives. METHODS: We evaluated 302 expertly validated SADS cases with suitable DNA (median age: 24 years; 65% males) who underwent next-generation sequencing using an extended panel of 77 primary electrical disorder and cardiomyopathy genes. Pathogenic and likely pathogenic variants were classified using American College of Medical Genetics (ACMG) consensus guidelines. The yield of combined molecular autopsy and clinical evaluation in 82 surviving families was evaluated. A gene-level rare variant association analysis was conducted in SADS cases versus controls. RESULTS: A clinically actionable pathogenic or likely pathogenic variant was identified in 40 of 302 cases (13%). The main etiologies established were catecholaminergic polymorphic ventricular tachycardia and long QT syndrome (17 [6%] and 11 [4%], respectively). Gene-based rare variants association analysis showed enrichment of rare predicted deleterious variants in RYR2 (p = 5 × 10(-5)). Combining molecular autopsy with clinical evaluation in surviving families increased diagnostic yield from 26% to 39%. CONCLUSIONS: Molecular autopsy for electrical disorder and cardiomyopathy genes, using ACMG guidelines for variant classification, identified a modest but realistic yield in SADS. Our data highlighted the predominant role of catecholaminergic polymorphic ventricular tachycardia and long QT syndrome, especially the RYR2 gene, as well as the minimal yield from other genes. Furthermore, we showed the enhanced utility of combined clinical and genetic evaluation

Plasma microparticle levels in polycystic ovary syndrome during treatment with metformin and oral contraceptives

Polycystic ovaries syndrome is the commonest endocrinopathy in women of reproductive age affecting their quality of life and health status, as it confers an increased cardiovascular risk. There are several therapeutic options for polycystic ovaries syndrome aiming at improving biochemical and hormonal parameters and preventing the complications. Routinely used therapeutic agents are combinations of contraceptives and metformin aiming at improvement of hyperandrogenemia and insulin resistance, respectively. Platelet microparticles (PMPs) are microvesicles shed from platelets following several stimuli and participate in inflammation, vascular homeostasis and thrombosis. They are involved in cardiovascular disease and were found increased in obese patients with polycystic ovaries syndrome. Agents that modulate hormonal aspects of PCOS could affect the level of PMPs. The aim of the present study was to evaluate the effects of oral contraceptive and metformin use for 6 and 12 months on platelet microparticles in normal-weight women with polycystic ovaries syndrome. Forty-five women with polycystic ovaries syndrome and 13 healthy women were recruited. All participants had normal body mass index (≤ 25 Kg/m2). Biochemical, hormonal and clinical parameters were recorded. Women with polycystic ovaries syndrome received treatment with combination of oral contraceptives, oral contraceptives+anti-androgens or metformin. Platelet microparticles were measured at baseline and after 6 and 12 months by means of flow cytometry. Statistical analysis was performed by means of the SPSS software (IBM SPSS Statistics for Windows, Version 25.0. Armonk, NY: IBM Corp., USA). At baseline, both groups had similar age (25±4 vs 26±2 years, p=0.064) and body mass index (23±3 vs 23±4 Kg/m2, p=0.830). Patients with polycystic ovaries syndrome had higher levels of platelet microparticles than controls (p=0.001) and increased after 6-month treatment with oral contraceptives (p=0.007). Subsequently, they decreased after 12-month treatment (p=0.046). Metformin had no effect on platelet microparticle levels. In detail, 8 patients received 3mg drospirenone+0,03mg ethinylestradiol, 20 patients received 0,035mg ethinylestradiol+2mg Cyproterone acetate, 5 patients received 0,035mg ethinylestradiol+2mg Cyproterone acetate+10mg Cyproterone acetate. Treatment for 6 and 12 months with combination of oral contraceptives improved FSH, LH, FAI και SHBG (p0.1). Finally, significant correlations between platelet microparticles and testosterone, DHEAS, AFI and insulin sensitivity indexes (insulin, glucose/insulin ratio, AUC-OGTT, HOMA-IR, QUICKI) were detected. The stronger correlations were observed with testosterone, DHEAS and FAI. In conclusion, platelet microparticle levels are increased in polycystic ovaries syndrome and further increase with oral contraceptive use. This effect could possibly contribute to the increased risk of venous thromboembolism associated with oral contraceptive use. However, further studies are needed to elucidate the exact role of platelet microparticles in polycystic ovaries syndrome.Το σύνδρομο των πολυκυστικών ωοθηκών αποτελεί τη συχνότερη ενδοκρινοπάθεια των γυναικών αναπαραγωγικής ηλικίας. Συχνότερα χρησιμοποιούμενες φαρμακευτικές αγωγές είναι τα από του στόματος συνδυασμένα αντισυλληπτικά δισκία και η μετφορμίνη, με στόχο την ομαλοποίηση της υπερανδρογοναιμίας και της αντίστασης στην ινσουλίνη, αντίστοιχα. Τα αιμοπεταλιακά μικροσωματίδια σχηματίζονται με εξωκυττάρωση από τα αιμοπετάλια και συμμετέχουν στη φλεγμονή, στη θρόμβωση και την ομοιοστασία των αγγείων. Στόχος της παρούσας μελέτης ήταν να διερευνηθεί εάν τα αιμοπεταλιακά μικροσωματίδια βελτιώνονται με τη χορήγηση συνδυασμένων αντισυλληπτικών ή μετφορμίνης για 6 και 12 μήνες. Πρόκειται για μία προοπτική τυχαιοποιημένη κλινική δοκιμή στην οποία συμμετείχαν άτομα με το σύνδρομο πολυκυστικών ωοθηκών και φυσιολογικό δείκτη μάζας σώματος (≤25Kg/m2) και υγιείς μάρτυρες. Συγκεντρώθηκαν στοιχεία σχετικά με το ατομικό αναμνηστικό, τη φυσική εξέταση, τα ανθρωπομετρικά στοιχεία, τις ορμονικές και βιοχημικές παραμέτρους. Επιπλέον, έγινε υπερηχογραφικός έλεγχος και προσδιορίσθηκαν τα αιμοπεταλιακά μικροσωματίδια. Ακολούθως, χορηγήθηκαν συνδυασμένα αντισυλληπτικά δισκία ή μετφορμίνη και μετρήθηκαν οι ίδιοι παράμετροι στους 6 και 12 μήνες. Τα αιμοπεταλιακά μικροσωματίδια προσδιορίστηκαν με την κυτταρομετρία ροής. Η στατιστική ανάλυση διενεργήθηκε με τη χρήση του λογισμικού SPSS (IBM SPSS Statistics for Windows, Version 25.0. Armonk, NY:IBM Corp., USA). Συμμετείχαν 45 γυναίκες με το σύνδρομο πολυκυστικών ωοθηκών και 13 υγιείς μάρτυρες. Οι δύο ομάδες είχαν παρόμοια ηλικία (25±4 vs 26±2 έτη, p=0,064) και δείκτη μάζας σώματος (23±3 vs 23±4 Kg/m2, p=0,830). Τα αιμοπεταλιακά μικροσωματίδια ήταν αυξημένα στην ομάδα των ατόμων με το σύνδρομο των πολυκυστικών ωοθηκών σε σύγκριση με τις υγιείς μάρτυρες (p=0,001). Οι γυναίκες με το σύνδρομο πολυκυστικών ωοθηκών έλαβαν είτε συνδυασμένο αντισυλληπτικό με ή χωρίς αντιανδρογόνο (ομάδα OCPs, n=33) είτε μετφορμίνη (n=12). Αναλυτικότερα, στην ομάδα που έλαβε συνδυασμένα αντισυλληπτικά χορηγήθηκαν: 1. 3mg drospirenone+0,03 mg ethinylestradiol (n=8), 2. 0,035 mg ethinylestradiol+2 mg Cyproteroneacetate (n=20), 3. 0,035 mg ethinylestradiol+2 mg Cyproteroneacetate+10 mg Cyproteroneacetate (n=5). Τα αιμοπεταλιακά μικροσωματίδια αυξήθηκαν με τη χορήγηση αντισυλληπτικών στους 6 μήνες (p=0,007) και ακολούθως μειώθηκαν στους 12 μήνες (p=0,046). Η μετφορμίνη δεν είχε καμία επίδραση στα επίπεδα των αιμοπεταλιακών μικροσωματιδίων (p>0,05). Η χορήγηση συνδυασμένων αντισυλληπτικών για 6 και 12 μήνες οδήγησε σε στατιστικά σημαντική μείωση των FSH, LH, FAI και SHBG (p<0,019). Οι ορμόνες: τεστοστερόνη και Δ4-A μεταβλήθηκαν σημαντικά μόνο μετά από αγωγή 12 μηνών (p<0,048). Η χορήγηση της μετφορμίνης στις γυναίκες με το σύνδρομο οδήγησε σε στατιστικά σημαντική διαφορά στις παραμέτρους LH, FAI και SHBG, FSH, στην Δ4-A, τεστοστερόνη και στην 17α-OHP (p<0,047). Διαπιστώθηκε στατιστικά σημαντική συσχέτιση μεταξύ των αιμοπεταλιακών μικροσωματιδίων και της τεστοστερόνης, της DHEAS, του δείκτη FAI και των δεικτών ινσουλινοαντίστασης (ινσουλίνη, λόγος γλυκόζης/ινσουλίνης, AUC-OGTT, HOMA-IR, QUICKI) εκ των οποίων οι ισχυρότερες συσχετίσεις ήταν μεταξύ των αιμοπεταλιακών μικροσωματιδίων και των ορμονικών δεικτών όπως η τεστοστερόνη, η DHEAS και του δείκτη FAI. Συμπερασματικά, τα αιμοπεταλιακά μικροσωματίδια είναι αυξημένα στα άτομα με το σύνδρομο των πολυκυστικών ωοθηκών συμβάλλοντας ενδεχομένως στον αυξημένο καρδιαγγειακό κίνδυνο. Η χορήγηση συνδυασμένων αντισυλληπτικών δισκίων αυξάνει βραχυπρόθεσμα τα αιμοπεταλιακά μικροσωματίδια και στη συνέχεια τα μειώνει συμβάλλοντας πιθανόν στον αυξημένο κίνδυνο φλεβικής θρομβοεμβολικής νόσου κατά τους πρώτους μήνες θεραπείας με αντισυλληπτικά. Περαιτέρω μελέτες απαιτούνται για να καθοριστεί ο παθογενετικός μηχανισμός που θα υποστηρίξει αυτή την υπόθεση

A Simple Food-Diverting Operation for Type 2 Diabetes Treatment. Preliminary Results in Humans with BMI 28-32 kg/m(2)

The feasibility of a simple side-to-side jejunoileal anastomosis (SJA) in non-morbidly obese individuals to control type 2 diabetes mellitus (T2DM) was studied in six diabetic patients with BMI 28-32. This novel procedure was performed in two Academic Centers and preliminary data is presented

The influence of combined oral contraceptives containing drospirenone on hypothalamic-pituitary-adrenocortical axis activity and glucocorticoid receptor expression and function in women with polycystic ovary syndrome

OBJECTIVE: Most women with PCOS have increased adrenal androgen production, enhanced peripheral metabolism of cortisol and elevation in urinary excretion of its metabolites. Increased cortisol clearance in PCOS is followed by a compensatory overdrive of the hypothalamic-pituitary-adrenocortical (HPA) axis. We hypothesized that oral contraceptives containing ethinylestradiol and drospirenone (EE-DRSP) could modulate glucocorticoid receptor (GR) expression and function and thus affect HPA axis activity in PCOS patients. DESIGN: We analyzed 12 women with PCOS (age 24.17 +/- 4.88 years; body mass index 22.05 +/- 3.97 kg/m(2)) treated for 12 months with EE-DRSP and 20 BMI matched controls. In all subjects testosterone, dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), cortisol (basal and after dexamethasone), concentrations of GR protein, phospo-GR211 protein, number of GR per cell (B-max) and its equilibrium dissociation constant (K-D) were measured. RESULTS: Before treatment, increased concentrations of testosterone and DHEAS (p<0.001, respectively), unaltered basal cortisol and an increased sensitivity (p<0.05) of the HPA axis to dexamethasone were observed in PCOS women in comparison to controls. After treatment, testosterone (p<0.01), DHEAS (p<0.05) and cortisol suppression after dexamethasone (p<0.01) were decreased in PCOS women. There were no changes in GR protein concentration, GR phosphorylation nor in the receptor functional parameters B-max and K-D in women with PCOS before and after the therapy, and in comparison to controls. CONCLUSIONS: Prolonged treatment with EE-DRSP in PCOS women decreased serum androgens and increased cortisol in the presence of decreased sensitivity of the HPA axis and did not exert changes in GR expression and function.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}11141009

The influence of combined oral contraceptives containing drospirenone on hypothalamic-pituitary-adrenocortical axis activity and glucocorticoid receptor expression and function in women with polycystic ovary syndrome

OBJECTIVE: Most women with PCOS have increased adrenal androgen production, enhanced peripheral metabolism of cortisol and elevation in urinary excretion of its metabolites. Increased cortisol clearance in PCOS is followed by a compensatory overdrive of the hypothalamic-pituitary-adrenocortical (HPA) axis. We hypothesized that oral contraceptives containing ethinylestradiol and drospirenone (EE-DRSP) could modulate glucocorticoid receptor (GR) expression and function and thus affect HPA axis activity in PCOS patients. DESIGN: We analyzed 12 women with PCOS (age 24.17 +/- 4.88 years; body mass index 22.05 +/- 3.97 kg/m(2)) treated for 12 months with EE-DRSP and 20 BMI matched controls. In all subjects testosterone, dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), cortisol (basal and after dexamethasone), concentrations of GR protein, phospo-GR211 protein, number of GR per cell (B-max) and its equilibrium dissociation constant (K-D) were measured. RESULTS: Before treatment, increased concentrations of testosterone and DHEAS (p<0.001, respectively), unaltered basal cortisol and an increased sensitivity (p<0.05) of the HPA axis to dexamethasone were observed in PCOS women in comparison to controls. After treatment, testosterone (p<0.01), DHEAS (p<0.05) and cortisol suppression after dexamethasone (p<0.01) were decreased in PCOS women. There were no changes in GR protein concentration, GR phosphorylation nor in the receptor functional parameters B-max and K-D in women with PCOS before and after the therapy, and in comparison to controls. CONCLUSIONS: Prolonged treatment with EE-DRSP in PCOS women decreased serum androgens and increased cortisol in the presence of decreased sensitivity of the HPA axis and did not exert changes in GR expression and function.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}11141009

Electrocardiographic differentiation between 'benign T-wave inversion' and arrhythmogenic right ventricular cardiomyopathy

AIMS: To characterize the most common electrocardiographic (ECG) abnormalities in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC), including anterior T-wave inversion (TWI) and to compare the characteristics of TWI in patients with ARVC and in a cohort of young healthy athletes and sedentary individuals. METHODS AND RESULTS : The study population consisted of 162 patients with a definite diagnosis of ARVC and 129 young controls with anterior TWI. Cardiac disease was excluded in all controls after a comprehensive diagnostic work-up. The ECG was abnormal in 131 patients with ARVC (81%). Abnormalities included anterior TWI (n\u2009=\u200982, 51%), QRS duration ratio V2:V5 >1.2 (n\u2009=\u200951, 31%), prolonged terminal S wave activation duration in V2 >55 ms (n\u2009=\u200942, 26%), inferior TWI (n\u2009=\u200930, 18%), and lateral TWI (n\u2009=\u200926, 16%). The J-point preceding anterior TWI was \u20091.2 (52%) and inferior or lateral TWI (47%). CONCLUSION: The ECG is frequently abnormal in patients with ARVC and anterior TWI is the most common feature. Anterior TWI is usually accompanied by other abnormalities in ARVC, which are uncommon in healthy individuals. J point <0.1\u2009mV preceding anterior TWI is not specific to ARVC and is observed in the majority of healthy individuals, including athletes, indicating a limited role for differentiating physiology or normal variants from ARVC