N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells

The cellular prion protein (PrPC) plays a fundamental role in prion disease. PrPC is a glycosylphosphatidylinositol (GPI)-anchored protein with two variably occupied N-glycosylation sites. In general, GPI-anchor and N-glycosylation direct proteins to apical membranes in polarized cells whereas the majority of mouse PrPC is found in basolateral membranes in polarized Madin-Darby canine kidney (MDCK) cells. In this study we have mutated the first, the second, and both N-glycosylation sites of PrPC and also replaced the GPI-anchor of PrPC by the Thy-1 GPI-anchor in order to investigate the role of these signals in sorting of PrPC in MDCK cells. Cell surface biotinylation experiments and confocal microscopy showed that lack of one N-linked oligosaccharide leads to loss of polarized sorting of PrPC. Exchange of the PrPC GPI-anchor for the one of Thy-1 redirects PrPC to the apical membrane. In conclusion, both N-glycosylation and GPI-anchor act on polarized sorting of PrPC, with the GPI-anchor being dominant over N-glycans

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

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Tuning the mechanical and morphological properties of self-assembled peptide hydrogels via control over the gelation mechanism through regulation of ionic strength and the rate of pH change

Hydrogels formed by the self-assembly of peptides are promising biomaterials. The bioactive and biocompatible molecule Fmoc-FRGDF has been shown to be an efficient hydrogelator via a &pi;-&beta; self-assembly mechanism. Herein, we show that the mechanical properties and morphology of Fmoc-FRGDF hydrogels can be effectively and easily manipulated by tuning both the final ionic strength and the rate of pH change. The increase of ionic strength, and consequent increase in rate of gelation and stiffness, does not interfere with the underlying &pi;-&beta; assembly of this Fmoc-protected peptide. However, by tuning the changing rate of the system\u27s pH through the use of glucono-&delta;-lactone to form a hydrogel, as opposed to the previously reported HCl methodology, the morphology (nano- and microscale) of the scaffold can be manipulated

Comparison between pulsed and continuous radiofrequency delivery

Das paroxysmale Vorhofflimmern (AF) wird zu 94% aus Foci aus dem Bereich von 2 bis 4 cm innerhalb der Pulmonalvenen getriggert [20]. Ein Ziel der Studie war die Klärung der Frage, inwieweit die Lokalisation der Ablationsnarbe im Bereich der Pulmonalvenen für einen Ablationserfolg ausschlaggebend ist. Als weiteres Studienziel galt es, die kontinuierliche Radiofrequenz-(RF)-Katheterablation mit der gepulsten Katheterablation zu vergleichen. Im Tierexperiment wurden Schweine lege artis anästhesiert und intubiert beatmet. Für jede Pulmonalvene wurden drei verschiedene anatomische Lokalisationen randomisiert ausgewählt: intraatrial, am Ostium der Pulmonalvene und innerhalb der Pulmonalvene selbst. Für die Ablation wurden drei verschiedene Energieeinstellungen verwendet: 30, 40 und 50 Watt (bei 55º C als Voreinstellung). Die Ablationen wurden mit einem zirkumferentiellen Ablationskatheter durchgeführt. Die Energiezufuhr zum Katheter wurde gepulst oder kontinuierlich gewählt. Die gepulste Katheterablation war bezüglich des Zeitmanagements sowie der Qualität der Homogenität und Transmuralität der kontinuierlichen Katheterablation überlegen. Bei der gepulsten Katheterablation traten weniger Komplikationen (PE, VT) auf als bei der kontinuierlichen Katheterablation. Die ostiale Pulmonalvenenisolation erwies sich als beste anatomische Lokalisation. Im Gegensatz dazu wiesen die intraatrialen Läsionen eine inhomogenere und eine weniger transmurale Ablation auf. Die Ablation innerhalb der Pulmonalvenen war signifikant häufiger mit einer Pulmonalvenenstenose (> 50% des Durchmessers) assoziiert. Die tierexperimentelle AF-Katheterablation im Bereich des Ostiums der Pulmonalvenen erzielte in Bezug auf die Homogenität der Ablationsnarbe und der Transmuralität der Läsion die besten Resultate. Des Weiteren wies die Ablation im Bereich des Ostiums ein geringeres Risiko hinsichtlich der Komplikation für eine Pulmonalvenenstenose auf. Die gepulste Katheterablation kann in signifikant kürzerer Zeit durchgeführt werden als die kontinuierliche Katheterablation. Bezüglich der Transmuralität und Homogenität der Läsionen konnte mit der gepulsten Katheterablation ein besseres Ergebnis erzielt werden als mit der kontinuierlichen Katheterablation.Atrial fibrillation is characterized by uncoordinated atrial activation often with irregular and high ventricular frequencies. Catheter ablation of the pulmonary veins (PV) has revolutionized treatment for invasive treatment of atrial fibrillation. The PV are often a trigger for the development of atrial fibrillation [20]. The aim of our study was first to evaluate different anatomical sites for PV-isolation: intraartial, ostial of the pulmonary veins or in the pulmonary veins it selves. The exact target for ablation is still unknown. The second aim of the study was to compare continuous radiofrequency (RF) delivery with pulsed RF. The animal experiments were performed in 8 anaesthetized and ventilated pigs. For each pulmonary vein three different anatomical sites for RF catheter ablation were selected: left atrial, the ostia of the PV or in the PV. In addition three different energy settings were used: 30, 40 and 50 Watts (55°C temperature presetting). The ablations were performed with an octapolar circumferential ablation catheter, either with continuous RF-energy delivery or with pulsed RF-energy delivery. The pulsed RF-energy delivery revealed the best results regarding transmurality (yes/no) and homogeneity (scaled). The best results for PV-isolation was the ostial of the PV. Intraatrial lesions were less homogenous and less transmural. Intrapulmonary vein ablation was associated with significant (> 50% diameter) stenosis of the PV (5/32). Ostial ablation of the PV may have the best results regarding homogeneity and transmurality with a low risk of PV stenosis. Pulsed energy delivery revealed the fastest way to create linear circumferential ostial lesions

Profile of SDB cohort at baseline and follow-up.

<p>At follow-up 16 children originally diagnosed with PS, 11 with Mild OSA and 8 with MS OSA returned. Of these, 18 had resolved and 17 had ongoing SDB. Twelve children who had resolved received treatment. Eight children who were unresolved also received treatment.</p

Mean domain scores on CBCL (3A) and BRIEF (3B) at baseline (black bars) and follow-up (white bars) for the Control, Resolved and Unresolved groups.

<p>The dotted line represents the population mean. The solid lines represent the between group differences. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001.</p