The impact of global and local composition on the stability of Triple Helical DNA

It is common practise in antisense technology to view third strand binding to be controlled by the same principles which are found to determine the stability of the double helix. In contrast to this view based on a general consideration of the various forces contributing to the binding energy of the third strand it was proposed that the dominant contributions will originate from electrostatic interactions. These electrostatic contributions can be subdivided into sequence independent repulsive forces between the negatively charged backbones and into sequence dependent attractive forces between the positively charged protonated Hoogsteen cytosines and the backbone phosphates. The observable changes in the stability of triple helices should be a reflection of the number (global composition) and distribution (local composition) of cytosines in the third strand. To this aim two families of 38-mer oligonucleotides were synthesized, which have as a common design feature a linear array of 10 homopurine bases followed by 10 homopyrimidine bases as Watson & Crick complementary strand to the homopurine region and ending in a 10 homopyrimidine residue stretch which binds to the W&C helix via Hoogsteen base-pairing. This arrangement of homopurine and homopyrimidine sections with connecting pyrimidine linkers allows the formation of intramolecular triple helices of predetermined stoichiometry and strand orientation. Physical (UV-spectroscopy, CD-spectroscopy and fluorimetry) and biochemical techniques (P1-nuclease digestion) have been used to show that the oligonucleotides undergo a stepwise folding process from a random coil into a hairpin with 3'dangling tail and then into a intramolecular triple helix. This folding occurs as a function of pH and/or ionic strength. The effect of local and global composition on the stability of the three conformational transitions has been evaluated from a comparison of the melting temperatures and the behavior of the phase boundaries of the different oligonucleotides. As the result of this thesis the following general rules emerge: The stability of the third strand depends on the particular combination of sequence, pH and ionic strength. At physiological conditions (pH 7.1, 150 mM Na⁺) thymines and cytosines contribute equally to the stability (global effect) provided that the cytosines are spaced by more than one thymine. (local effect). Below pH 7.1 (150 mM Na⁺) the stability increases linearly with the number of cytosines and at pH above pH 7.1 ( 150 mM Na⁺) it decreases. At ionic strength below 400 mM Na⁺ (pH 6. 75) the stability increases with the number of cytosine while above 400 mM Na⁺ (pH 6. 75) it decreases. Based on these results a rational approach for the design of oligonucleotide third strands and the choice of appropriate environmental conditions for the formation of a particular triple helix becomes feasible

EKG und klinischer Verlauf vor und nach Implantation eines konventionellen Schrittmachers zur Evaluierung eines Kollektives mit verminderter Prognose:eine retrospektive Analyse an 298 Patienten der Schüchtermannklinik aus den Jahren 1997 bis 2000

Anhand klinischer,elektrokardiographischer und echokardiographischer Befunde untersuchten wir,welche Parameter sich auf die Prognose von Schrittmacherpatienten auswirken. Vor der Schrittmacherimplantation lag bei 75% der Patienten die QRS-Breite unter 120 msec,danach zeigten 77,2% der Patienten stimulationsbedingt eine QRS-Breite über 120 msec, bei 6,7% der Patienten war der stimulierte QRS-Komplex über 200 msec breit. Im Untersuchungszeitraum von 45 Monaten verstarben 14 von 177 nachbetreuten Patienten. In der multivariaten Analyse zeigten sich die Breite der stimulerten QRS-Komplexe über 200 ms und das präoperative NYHA-Stadium statistisch signifikant für die Ein-Jahres-Mortalität nach der Schrittmacherimplantation. Zusammenfassend kann das Ausmaß der stimulationsinduzierten QRS-Verbreiterung nach der Schrittmacherimplantation als Marker für ein erhöhtes Mortalitätsrisiko im ersten Jahr nach dem Eingriff angesehen werden

Schlussbericht zum InnoProfile Forschungsvorhaben Methoden und Baustoffe zur nutzerorientierten Bausanierung

Nutzerorientierte Bausanierung bedeutet eine gegenüber dem konventionellen Vorgehen deutlich verstärkte Ausrichtung des Planungs- und Sanierungsprozesses auf die Anforderungen und Bedürfnisse des zukünftigen Nutzers eines Gebäudes. Dies hat einerseits ein hochwertigeres Produkt zum Ergebnis, erfordert andererseits aber auch den Einsatz neuer Methoden und Baustoffe sowie ein vernetztes Zusammenarbeiten aller am Bauprozess Beteiligten. Der Fokus der Publikation liegt dabei auf den Bereichen, die eine hohe Relevanz für die nutzerorientierte Bausanierung aufweisen. Dabei handelt es sich insbesondere um: Computergestütztes Bauaufmaß und digitale Bauwerksmodellierung (BIM), bauphysikalische Methoden zur Optimierung von Energieeffizienz und Behaglichkeit bei der Sanierung von Bestandsgebäuden, zerstörungsfreie Untersuchungsmethoden im Rahmen einer substanzschonenden Bauzustandsanalyse und Entwicklung von Ergänzungsbaustoffen. Das Projekt nuBau ist eine Kooperation zwischen den Fakultäten Bauingenieurwesen und Architektur der Bauhaus-Universität Weimar. Die beteiligten Professuren sind: Bauphysik, Informatik in der Architektur, Polymere Werkstoffe und Werkstoffe des Bauens

Energy Metabolites as Biomarkers in Ischemic and Dilated Cardiomyopathy

With more than 25 million people affected, heart failure (HF) is a global threat. As energy production pathways are known to play a pivotal role in HF, we sought here to identify key metabolic changes in ischemic- and non-ischemic HF by using a multi-OMICS approach. Serum metabolites and mRNAseq and epigenetic DNA methylation profiles were analyzed from blood and left ventricular heart biopsy specimens of the same individuals. In total we collected serum from n = 82 patients with Dilated Cardiomyopathy (DCM) and n = 51 controls in the screening stage. We identified several metabolites involved in glycolysis and citric acid cycle to be elevated up to 5.7-fold in DCM (p = 1.7 × 10−6 ). Interestingly, cardiac mRNA and epigenetic changes of genes encoding rate-limiting enzymes of these pathways could also be found and validated in our second stage of metabolite assessment in n = 52 DCM, n = 39 ischemic HF and n = 57 controls. In conclusion, we identified a new set of metabolomic biomarkers for HF. We were able to identify underlying biological cascades that potentially represent suitable intervention targets

New genetic loci link adipose and insulin biology to body fat distribution.

Body fat distribution is a heritable trait and a well-established predictor of adverse metabolic outcomes, independent of overall adiposity. To increase our understanding of the genetic basis of body fat distribution and its molecular links to cardiometabolic traits, here we conduct genome-wide association meta-analyses of traits related to waist and hip circumferences in up to 224,459 individuals. We identify 49 loci (33 new) associated with waist-to-hip ratio adjusted for body mass index (BMI), and an additional 19 loci newly associated with related waist and hip circumference measures (P < 5 × 10(-8)). In total, 20 of the 49 waist-to-hip ratio adjusted for BMI loci show significant sexual dimorphism, 19 of which display a stronger effect in women. The identified loci were enriched for genes expressed in adipose tissue and for putative regulatory elements in adipocytes. Pathway analyses implicated adipogenesis, angiogenesis, transcriptional regulation and insulin resistance as processes affecting fat distribution, providing insight into potential pathophysiological mechanisms

Causal effect of plasminogen activator inhibitor type 1 on coronary heart disease

Background--Plasminogen activator inhibitor type 1 (PAI-1) plays an essential role in the fibrinolysis system and thrombosis. Population studies have reported that blood PAI-1 levels are associated with increased risk of coronary heart disease (CHD). However, it is unclear whether the association reflects a causal influence of PAI-1 on CHD risk. Methods and Results--To evaluate the association between PAI-1 and CHD, we applied a 3-step strategy. First, we investigated the observational association between PAI-1 and CHD incidence using a systematic review based on a literature search for PAI-1 and CHD studies. Second, we explored the causal association between PAI-1 and CHD using a Mendelian randomization approach using summary statistics from large genome-wide association studies. Finally, we explored the causal effect of PAI-1 on cardiovascular risk factors including metabolic and subclinical atherosclerosis measures. In the systematic meta-analysis, the highest quantile of blood PAI-1 level was associated with higher CHD risk comparing with the lowest quantile (odds ratio=2.17; 95% CI: 1.53, 3.07) in an age- and sex-adjusted model. The effect size was reduced in studies using a multivariable-adjusted model (odds ratio=1.46; 95% CI: 1.13, 1.88). The Mendelian randomization analyses suggested a causal effect of increased PAI-1 level on CHD risk (odds ratio=1.22 per unit increase of log-transformed PAI-1; 95% CI: 1.01, 1.47). In addition, we also detected a causal effect of PAI-1 on elevating blood glucose and high-density lipoprotein cholesterol. Conclusions--Our study indicates a causal effect of elevated PAI-1 level on CHD risk, which may be mediated by glucose dysfunction

Genome-wide association study of circulating interleukin 6 levels identifies novel loci

Interleukin 6 (IL-6) is a multifunctional cytokine with both pro- and anti-inflammatory properties with a heritability estimate of up to 61%. The circulating levels of IL-6 in blood have been associated with an increased risk of complex disease pathogenesis. We conducted a two-staged, discovery and replication meta genome-wide association study (GWAS) of circulating serum IL-6 levels comprising up to 67428 (ndiscovery=52654 and nreplication=14774) individuals of European ancestry. The inverse variance fixed effects based discovery meta-analysis, followed by replication led to the identification of two independent loci, IL1F10/IL1RN rs6734238 on chromosome (Chr) 2q14, (Pcombined=1.8x10-11), HLA-DRB1/DRB5 rs660895 on Chr6p21 (Pcombined=1.5x10-10) in the combined meta-analyses of all samples. We also replicated the IL6R rs4537545 locus on Chr1q21 (Pcombined=1.2x10-122). Our study identifies novel loci for circulating IL-6 levels uncovering new immunological and inflammatory pathways that may influence IL-6 pathobiology.</p

Development of a compact test facility for SRF Photoelectron injectors

SHF Photoelektroneninjektoren sind eine vielversprechende Elektronquelle für hochbrillante Teichenbeschleuniger mit hohem mittlerem Strom und kurzen Teilchenpulsen, wie FELs und ERLs. Für das ERL Projekt bERLinPro wurde einer unabhängige Testanlage GunLab entwickelt um die Leistungsfähigkeit der Injektoren und die Strahlparameter zu überprüfen. Darüber hinaus können neue Komponenten zur Strahldiagnose getestet werden. Die Hauptaufgabe von GunLab ist die Beschreibung des vollständigen sechsdimensionalen Phasenraums der Elektronen in Abhängigkeit aller Injektorparameter. Die Anlage besteht aus einer kompakten Diagnosestrahlführungan dem SHF Photoelektroneninjektoren und einem Kathodenlasersystem. Im Rahmen dieser Arbeit wurden analytische und numerische Studien zu den SHF Photoelektroneninjektoren durchgeführt, um zu erwartende Strahlparameter zu detektieren und die Anforderungen an die Strahldiagnose festzulegen. Darüber hinaus wurden verschiedene Emittanzbeiträge der einzelnen Injektorkomponenten untersucht. Diesbezüglich wurde das Magnetfeld des aktuellen Solenoiden kartiert und auf Asymmetrien getestet, die ebenfalls zu Emittanzvergrößerungen beitragen können. Eine der wesentlichen Komponenten der Diagnosestrahlführung ist das (transversale) Phasenraummesssystem, für das eine besondere Magnetgeometrie entwickelt wurde. Weitere Diagnose Komponenten sind ein optimierter Spektrometerdipol und eine transversal ablenkende Kavität, durch die sich zusammen mit zwei Quadrupolmagneten die Scheibenemittanz bestimmen lässt. Für GunLab wurden unterschiedliche optische Messsysteme entwickelt und optimiert. Der herausforderndste Aufbau ist dabei das Strahl-Halo Messsystem. Es ermöglicht die Beobachtung der transversalen Ladungsverteilung über einen Dynamikbereich von bis zu 6 Größenordnungen. Die Leistungsfähigkeit und die Auflösung aller Messsysteme und Messroutinen wurden bestimmt, um die Visualisierung des kompletten Phasenraums durch GunLab sicher zu stellen.SRF photoelectron injectors are promising electron sources for high brightness accelerators with high average current and short pulse duration like FELs and ERLs. For the ERL project bERLinPro an independent test facility called GunLab was developed and set up to optimize the operation performance of SRF photoinjectors and the electron beam parameters. Furthermore, GunLab allows to investigate the operation of different kinds of photocathodes in the environment of an SRF accelerator and to study new beam diagnostic concepts. Of outmost importance is the characterization of the full six dimensional phase space as a function of all injector parameters. GunLab consists of the compact diagnostic beam line, connected to the SRF photoinjetor module, and a drive laser. In the context of this thesis, analytical and numerical investigations of the SRF photoinjector were performed to estimate beam parameter ranges and to determine the diagnostics requirements. Furthermore, various emittance contributors of the injector were determined. Thereby the magnetic field of the final designed solenoid was measured to determine field asymmetries, which are one major source of emittance growth. One of the central diagnostic components of the beamline is the (horizontal) phase space scanner system. For this purpose, a dedicated air-coil magnet design was developed. Additional diagnostic components include an optimized spectrometer system, a transverse deflecting cavity (TCav) and two quadrupole magnets, to determine longitudinal and sliced emittance. For GunLab different optical measurement systems were developed and optimized, the most challenging setup is a beam halo measurement system. This device is able to observe the transverse charge density with a dynamic range of up to 6 orders of magnitude. The performance and the resolution of all measurement systems and routines for GunLab were determined to ensure the visualization of the electron beam phase space