Two views of native alpha 2-macroglobulin are revealed by electron microscopy of negatively stained samples; in one view the molecule resembles a padlock and in the other, a pair of lips. Interconversion of the two views upon tilting establishes that these are two different projected views of the same structure. Furthermore, the two views are related by a 45 degrees rotation about their major a...

Labeling of a covalently closed circular double-stranded DNA was achieved using a so-called 'padlock oligonucleotide'. The oligonucleotide was targeted to a sequence which is present in the replication origin of phage f1 and thus in numerous commonly used plasmids. After winding around the double-stranded target DNA sequence by ligand-induced triple helix formation, a biotinylated oligonucleoti...

Rare tumor-specific mutations in patient samples serve as excellent markers to monitor the course of malignant disease and responses therapy clinical routine, improved assay techniques are needed for broad adoption. We describe herein - superRCA assays which provides rapid highly specific detection DNA sequence variants present at very low frequencies samples. Using a standard flow cytometer we...

In situ padlock/rolling circle amplification (RCA) is a method used to amplify, visualize, and quantify target DNA molecules in cells. However, the multiple reaction steps involved make this technique costly and cumbersome. We developed a novel, simplified, automated microfluidic system for RCA, and demonstrated its effectiveness by counting amplified mitochondrial DNA fragments in HeLa cells. ...

Background: The ultrasonography probes are cleaned by absorbent soft, dry cloth. This question arose whether linear and convex ultrasound probes coupling with gel could perform as a means for nosocomial infections transmission, and which method is economical and more efficient for cleaning the probes. This study was conducted to answer these questions. Methods: One hundred – ninety two patients...

Highly specific high-throughput assays will be required to take full advantage of the accumulating information about the macromolecular composition of cells and tissues, in order to characterize biological systems in health and disease. We discuss the general problem of detection specificity and present the approach our group has taken, involving the reformatting of analogue biological informat...

BACKGROUND We developed a non-survival in vivo model for testing of gastric natural orifice translumenal endoscopic surgery (NOTES) closures based on the gastric yield pressure. The aim of this study was to test our model comparing different endoscopic closure techniques with surgical closure of a NOTES gastric incision. METHODS Laparotomy was performed in 30 pigs. One tube for air inflation ...

In this thesis, we tried to improve the initiation of Rolling Circle Amplification (RCA) on genomic DNA using an approach known as Single Primer Extension (SPE). Usually, a synthetic DNA target provides a high signal on RCA after padlock probe hybridisation due to its short 3’ end. However, unfragmented genomic DNA provides low efficiency for RCA initiation due to its long 3’ end. During SPE, t...