Simon Schwill

MicroRNAs Differentially Expressed in Postnatal Aortic Development Downregulate Elastin via 3′ UTR and Coding-Sequence Binding Sites

Elastin production is characteristically turned off during the maturation of elastin-rich organs such as the aorta. MicroRNAs (miRNAs) are small regulatory RNAs that down-regulate target mRNAs by binding to miRNA regulatory elements (MREs) typically located in the 3′ UTR. Here we show a striking up-regulation of miR-29 and miR-15 family miRNAs during murine aortic development with commensurate down-regulation of targets including elastin and other extracellular matrix (ECM) genes. There were a total of 14 MREs for miR-29 in the coding sequences (CDS) and 3′ UTR of elastin, which was highly significant, and up to 22 miR-29 MREs were found in the CDS of multiple ECM genes including several collagens. This overrepresentation was conserved throughout mammalian evolution. Luciferase reporter assays showed synergistic effects of miR-29 and miR-15 family miRNAs on 3′ UTR and coding-sequence elastin constructs. Our results demonstrate that multiple miR-29 and miR-15 family MREs are characteristic for some ECM genes and suggest that miR-29 and miR-15 family miRNAs are involved in the down-regulation of elastin in the adult aorta.

Predictive Risk Factors for Patients With Cirrhosis Undergoing Heart Surgery

BACKGROUND Empiric experiences suggest higher mortality and complication risk for patients with cirrhosis of the liver after cardiac surgery. However, cirrhosis is not considered a risk factor in either the EuroSCORE or The Society of Thoracic Surgeons score. We report a large single-center experience of patients with cirrhosis undergoing cardiac surgery with extracorporeal circulation and aimed to evaluate the severity of cirrhosis as a predictor of outcome. METHODS During 2001 and 2011, we operated on 109 consecutive patients (average age, 64 years; 82 male) diagnosed for cirrhosis with cardiopulmonary bypass for different indications. Thirty-day mortality and long-term mortality were set as primary study end points. RESULTS Thirty-day mortality was 26%, and 5-year survival was 19%. Patients categorized as Child-Turcotte-Pugh (CHILD) C (n=6; 67% 30-day survival; 0% 5-year survival) and B (n=30; 60%; 5%) had worse 30-day and 5-year survival compared with patients categorized as CHILD A (n=73; 80%; 25%). For 30-day mortality, preoperative EuroSCORE (p=0.015), model for end-stage liver disease (MELD) score (p=0.006), albumin (p=0.023), total protein (p=0.01), and myocardial infarction (p=0.049) revealed significant differences between survivors and nonsurvivors. Multivariate logistic regression identified only MELD score (odds ratio [OR], 1.12; 95% confidence interval [CI], 1.03 to 1.23; p=0.011) and total protein (OR, 0.97; 95% CI, 0.95 to 1; p=0.049) were connected with increased 30-day mortality. Cox regression analysis revealed EuroSCORE (OR, 1.02; 95% CI, 1.01 to 1.03; p<0.0001) and MELD (OR, 1.06; 95% CI, 1.01 to 1.12; p=0.016) predicting the overall mortality. Receiver operating characteristic analysis indicated significant predictive power of MELD (p=0.001) and EuroSCORE (p=0.027) for 30-day mortality. CONCLUSIONS Patients with cirrhosis undergoing heart surgery with extracorporeal circulation have a poor prognosis. Several preoperative factors are related to outcome. EuroSCORE and MELD score may help to evaluate operation risk and indication.

An alternative surgical approach for the combined treatment of pectus excavatum and acute aortic dissection type-A in Marfan syndrome

Acute aortic dissection type-A (AADA) is a life-threatening condition especially in patients with Marfan syndrome (MFS) simultaneously suffering from severe pectus excavatum (PE). We report on emergency surgery for combined treatment of PE and AADA in a patient with MFS using an alternative approach. It leads to excellent exposure of the dislocated heart and great vessels enabling Bentall procedure followed by funnel chest repair with modified technique of Adkins and Blades. We achieved favorable functional and cosmetic results. Therefore, we conclude the surgical approach presented is feasible for standard treatment of AADA and consecutive repair of PE.

Loss of Endothelial Barrier in Marfan Mice (mgR/mgR) Results in Severe Inflammation after Adenoviral Gene Therapy

Objectives Marfan syndrome is an autosomal dominant inherited disorder of connective tissue. The vascular complications of Marfan syndrome have the biggest impact on life expectancy. The aorta of Marfan patients reveals degradation of elastin layers caused by increased proteolytic activity of matrix metalloproteinases (MMPs). In this study we performed adenoviral gene transfer of human tissue inhibitor of matrix metalloproteinases-1 (hTIMP-1) in aortic grafts of fibrillin-1 deficient Marfan mice (mgR/mgR) in order to reduce elastolysis. Methods We performed heterotopic infrarenal transplantation of the thoracic aorta in female mice (n = 7 per group). Before implantation, mgR/mgR and wild-type aortas (WT, C57BL/6) were transduced ex vivo with an adenoviral vector coding for human TIMP-1 (Ad.hTIMP-1) or β-galactosidase (Ad.β-Gal). As control mgR/mgR and wild-type aortas received no gene therapy. Thirty days after surgery, overexpression of the transgene was assessed by immunohistochemistry (IHC) and collagen in situ zymography. Histologic staining was performed to investigate inflammation, the neointimal index (NI), and elastin breaks. Endothelial barrier function of native not virus-exposed aortas was evaluated by perfusion of fluorescent albumin and examinations of virus-exposed tissue were performed by transmission electron microscopy (TEM). Results IHC and ISZ revealed sufficient expression of the transgene. Severe cellular inflammation and intima hyperplasia were seen only in adenovirus treated mgR/mgR aortas (Ad.β-Gal, Ad.hTIMP-1 NI: 0.23; 0.43), but not in native and Ad.hTIMP-1 treated WT (NI: 0.01; 0.00). Compared to native mgR/mgR and Ad.hTIMP-1 treated WT aorta, the NI is highly significant greater in Ad.hTIMP-1 transduced mgR/mgR aorta (p = 0.001; p = 0.001). As expected, untreated Marfan grafts showed significant more elastolysis compared to WT (p = 0.001). However, elastolysis in Marfan aortas was not reduced by adenoviral overexpression of hTIMP-1 (compared to untreated Marfan aorta: Ad.hTIMP-1 p = 0.902; control Ad.β-Gal. p = 0.165). The virus-untreated and not transplanted mgR/mgR aorta revealed a significant increase of albumin diffusion through the endothelial barrier (p = 0.037). TEM analysis of adenovirus-exposed mgR/mgR aortas displayed disruption of the basement membrane and basolateral space. Conclusions Murine Marfan aortic grafts developed severe inflammation after adenoviral contact. We demonstrated that fibrillin-1 deficiency is associated with relevant dysfunction of the endothelial barrier that enables adenovirus to induce vessel-harming inflammation. Endothelial dysfunction may play a pivotal role in the development of the vascular phenotype of Marfan syndrome.

Die Maus als Modell für die Grundlagenforschung bei Marfan-Syndrom

ZusammenfassungDas Marfan-Syndrom ist mit einer Inzidenz von 2–3 Patienten auf 10.000 Geburten eine seltene hereditäre, systemische Bindegewebsstörung. Marfan-Patienten sind durch die Ausbildung von Aneurysmen der Aorta und durch spontane Aortendissektionen gefährdet. Das primäre Ziel bei der Therapie ist die Prävention von lebensbedrohlichen kardiovaskulären Ereignissen. Die Pathogenese der Aortenerkrankung ist nicht abschließend geklärt. Für die Entwicklung neuer Therapieansätze ist eine detaillierte Erforschung der Pathophysiologie vor allem auf molekularer Ebene notwendig. Aus ethischen Überlegungen ist für die Entwicklung experimenteller Therapieansätze ein präklinisches Modell sinnvoll. Hauptanspruch an das Modell ist die Imitation der humanen Pathologie für eine möglichst genaue Translation von Modell zu Mensch. Der vorliegende Beitrag stellt ein Mausmodell des Marfan-Syndroms vor. Die mgR/mgR-Maus zeigt auf makroskopischer und mikroskopischer Ebene phänotypische Manifestationen des Marfan-Syndroms. Damit ermöglicht die mgR/mgR-Maus ein besseres Verständnis der Pathophysiologie und die Entwicklung neuer Therapieansätze zur Behandlung der kardiovaskulären Manifestationen des Marfan-Syndroms.AbstractMarfan syndrome is a rare hereditary systemic connective tissue disease with a prevalence of 2–3 in 10,000 births. Aneurysm formation and aortic dissection still remain the most life-threatening complications in Marfan syndrome but the pathogenesis of the aortic disease is poorly understood. The primary goal of treatment is the prevention of life-threatening cardiovascular events. Further investigations on the molecular mechanisms of aortic disease are crucial for the development of new treatment-strategies. Experimental therapies should be established in a preclinical model which needs to mimic the human pathology of Marfan syndrome for a valuable translation from model to human. This report provides information on a murine model of Marfan syndrome. The mgR/mgR mouse presents with phenotypic manifestations of Marfan syndrome at the macroscopic and microscopic levels. Further investigations on mgR/mgR mice could reveal new insights into the molecular understanding of aortic disease and could offer targets to improve treatment of the life-threatening cardiovascular manifestations of Marfan syndrome.

Komplementäre und Integrative Medizin in der Facharztweiterbildung Allgemeinmedizin: Ergebnisse einer Bedarfserhebung bei Ärzten in Weiterbildung

Hintergrund: Inhalte aus den Bereichen der klassischen Naturheilverfahren und Komplementärmedizin sind im Rahmen der ärztlichen Approbationsordnung curricular verankert. Im Gegensatz dazu werden in den Weiterbildungsordnungen zum Facharzt für Allgemeinmedizin keine komplementärmedizinischen Inhalte abgebildet. Bisher ist nicht bekannt, ob Ärzte in Weiterbildung (ÄiW) zum Facharzt für Allgemeinmedizin innerhalb ihrer Weiterbildung mit komplementärmedizinischen Verfahren in Kontakt kommen und ob sie solche erlernen bzw. erlernen möchten. Das Ziel der vorliegenden Studie war die Erhebung der Einstellung zum und des Weiterbildungsbedarfs im Bereich Komplementärmedizin bei ÄiW zum Facharzt für Allgemeinmedizin. Methoden: In Rahmen einer Querschnittsstudie erfolgte eine Fragebogen-gestützte Umfrage unter ÄiW zum Facharzt für Allgemeinmedizin innerhalb des Weiterbildungsprogramms «Verbundweiterbildungplus Baden-Württemberg». Diese wurde onlinebasiert initiiert und durch eine papierbasierte Umfrage komplettiert. Ergebnisse: Insgesamt nahmen 138 Teilnehmer der Verbundweiterbildungplus Baden-Württemberg an der Umfrage teil. Der Gesamtrücklauf betrug damit 28%. Dabei zeigte sich, dass die Teilnehmenden ein hohes Interesse an Komplementärmedizin hatten. Gleichzeitig gaben sie an, Unsicherheiten sowohl inhaltlicher als auch formaler Art wahrzunehmen (z.B. Evidenz der einzelnen Methoden und Kostenübernahme durch gesetzliche Krankenkassen). Die große Mehrheit der befragten Ärzte befürwortete, dass in der Weiterbildung zum Facharzt Allgemeinmedizin Kompetenzen aus dem Bereich Komplementärmedizin vermittelt werden. Schlussfolgerungen: Vor dem Hintergrund der weiten Verbreitung komplementärmedizinischer Methoden im hausärztlichen Alltag muss diskutiert werden, ob im Rahmen der Weiterbildung zum Facharzt für Allgemeinmedizin Basiskompetenzen für diesen Bereich definiert werden sollten. Diese könnten beispielsweise in das «Kompetenzbasierte Curriculum Allgemeinmedizin» der Deutschen Gesellschaft für Allgemeinmedizin und Familienmedizin (DEGAM) einfließen.

Novel oral anticoagulants in primary care in patients with atrial fibrillation: a cross-sectional comparison before and after their introduction

BackgroundNovel oral anticoagulation (NOAC) has been introduced in recent years, but data on use in atrial fibrillation (AF) in primary care setting is scarce. In Germany, General Practitioners are free to choose type of oral anticoagulation (OAC) in AF. Our aim was to explore changes in prescription-rates of OAC in German primary care before and after introduction of NOAC on the market.MethodsData of a representative morbidity registration project in primary care in Germany (CONTENT) were analysed. Patients with AF in 2011 or 2014 were included (before and after broad market authorization of NOAC, respectively). We defined three independent groups: patients from 2011 without follow-up (group A), patients from 2014 but without previous record in 2011 (group B) and patients with AF and records in 2011 and 2014 (group C).Results2642 patients were included. Group A (n = 804) and B (n = 755) were comparable regarding patient characteristics. 87.3% of group A and 84.8% of group B had CHA2DS2-VASc-Score ≥ 2, indicating a need for oral anticoagulation (OAC). Prescription of OAC increased from 23.1% (n = 186) to 42.8% (n = 323, p < .01) with stable use of vitamin-k-antagonist (22.6–24.9%). NOAC increased from 0.6 to 19.2% (p < .01). Monotherapy with Acetylsalicylic acid (ASA) decreased from 15.3% (n = 123) to 8.2% (n = 62, p < .01). In group C (n = 1083), OAC increased from 35.3 to 55.4% (p < .01), with stable prescription rate of vitamin-k-antagonist (34.4–35.7%). NOAC increased from 0.9 to 21.5% (p < .01).ConclusionsIn summary, our study showed a significant increase of OAC over time, which is fostered by the use of NOAC but with a stable rate of VKA and a sharp decrease of ASA. Patients on VKA are rarely switched to NOAC, but new patients with AF are more likely to receive NOAC.

AP-1 Oligodeoxynucleotides Reduce Aortic Elastolysis in a Murine Model of Marfan Syndrome

Marfan syndrome is characterized by high expression of matrix metalloproteinases (MMPs) in aortic smooth muscle cells (AoSMCs) associated with medial elastolysis and aortic root aneurysm. We aimed to reduce aortic elastolysis through decrease of MMP expression with decoy oligodeoxynucleotides (dODNs) neutralizing the transcription factor activating factor-1 (AP-1). AP-1 abundance in nuclear extracts as well as MMP-2 and MMP-9 expression were significantly increased in isolated mAoSMC of mgR/mgR Marfan mice compared to wild-type cells. Exposure to AP-1 neutralizing dODNs resulted in a significant reduction of basal and interleukin-1β-stimulated MMP expression and activity in mAoSMCs. Moreover, increased migration and formation of superoxide radical anions was substantially decreased in mAoSMCs by AP-1 dODN treatment. Aortic grafts from donor Marfan mice were treated with AP-1- dODN ex vivo and implanted as infrarenal aortic interposition grafts in mgR/mgR mice. Pretreatment of aortic grafts with AP-1 dODN led to reduced elastolysis, macrophage infiltration, and MMP activity. Permeability of the endothelial monolayer was increased for dODN in mgR/mgR aortae with observed loss of tight junction proteins ZO-1 and occludin, enabling dODN to reach the tunica media. Targeting AP-1 activity offers a new potential strategy to treat the vascular phenotype associated with Marfan syndrome.