Molecular characterization of two new alternaviruses identified in members of the fungal family Nectriaceae.

Since the first report in 2009, at least ten additional viruses have been identified and assigned to the proposed virus family Alternaviridae. Here we report two new mycoviruses tentatively assigned to this family, both identified as members of the fungal family Nectriaceae, which were isolated from surface-disinfected apple roots (Malus x domestica, Borkh.) affected by apple replant disease (ARD). ARD is a highly complex, worldwide-occurring disease resulting from plant reactions to a disturbed (micro)-biome and leads to high economic losses every year. The first alternavirus characterized in this study was identified in a Dactylonectria torresensis isolate. The virus was tentatively named dactylonectria torresensis alternavirus 1 (DtAV1) as the first member of the proposed new species Alternavirus dactylonectriae. The second virus was identified in an isolate of Ilyonectria robusta and was tentatively named ilyonectria robusta alternavirus 1 (IrAV1) as the first member of the proposed new species Alternavirus ilyonectriae. Full genomic sequences of the viruses were determined and are presented. Further, we found hints for putative components of a methyl transferase machinery using in silico approaches. This putative protein domain is encoded by segment 2. However, this result only establishes the basis for subsequent studies in which the function must be confirmed experimentally in vitro. Thus, this is the first study where a function is predicted to all three genomic segments within the group of the alternaviruses. These findings provide further insights into the virome of ARD-associated fungi and are therefore another brick in the wall of understanding the complexity of the disease.

Enhancing the stability of DNA origami nanostructures: staple strand redesign versus enzymatic ligation.

DNA origami structures have developed into versatile tools in molecular sciences and nanotechnology. Currently, however, many potential applications are hindered by their poor stability, especially under denaturing conditions. Here we present and evaluate two simple approaches to enhance DNA origami stability. In the first approach, we elevated the melting temperature of nine critical staple strands by merging the oligonucleotides with adjacent sequences. In the second approach, we increased the global stability by enzymatically ligating all accessible staple strand ends directly. By monitoring the gradual urea-induced denaturation of a prototype triangular DNA origami modified by these approaches using atomic force microscopy, we show that rational redesign of a few, critical staple strands leads to a considerable increase in overall stability at high denaturant concentration and elevated temperatures. In addition, enzymatic ligation yields DNA nanostructures with superior stability at up to 37 °C and in the presence of 6 M urea without impairing their shape. This bio-orthogonal approach is readily adaptable to other DNA origami structures without the need for synthetic nucleotide modifications when structural integrity under harsh conditions is required.

Dynamics of DNA Origami Lattice Formation at Solid-Liquid Interfaces.

The self-organized formation of regular patterns is not only a fascinating topic encountered in a multitude of natural and artificial systems, but also presents a versatile and powerful route toward large-scale nanostructure assembly and materials synthesis. The hierarchical, interface-assisted assembly of DNA origami nanostructures into regular, 2D lattices represents a particularly promising example, as the resulting lattices may exhibit an astonishing degree of order and can be further utilized as masks in molecular lithography. Here, we thus investigate the development of order in such 2D DNA origami lattices assembled on mica surfaces by employing in situ high-speed atomic force microscopy imaging. DNA origami lattice formation is found to resemble thin-film growth in several aspects. In particular, the Na+/Mg2+ ratio controls DNA origami adsorption, surface diffusion, and desorption, and is thus equivalent in its effects to substrate temperature which controls adatom dynamics in thin-film deposition. Consequently, we observe a pronounced dependence of lattice order on Na+ concentration. At low Na+ concentrations, lattice formation resembles random deposition and results in unordered monolayers, whereas very high Na+ concentrations are accompanied by rapid diffusion and especially DNA origami desorption, which prevent lattice formation. At intermediate Na+ concentrations, highly ordered DNA origami lattices are obtained that display an intricate symmetry, stemming from the complex shape of the employed Rothemund triangle. Nevertheless, even under such optimized conditions, the lattices display a considerable number of defects, including grain boundaries, point and line defects, and screw-like dislocations. By monitoring the dynamics of selected lattice defects, we identify mechanisms that limit the obtainable degree of lattice order. Possible routes toward further increasing lattice order by postassembly annealing are discussed.

Quadriceps force during knee extension after non-hinged and hinged TKA: an in vitro study.

BACKGROUND AND PURPOSE: Problems during knee extension, due to kinematic alterations, are not uncommon after total knee arthroplasty. Hinged prostheses provide higher stability than non-hinged designs and may minimize these alterations. Thus, in this in vitro study we investigated the quadriceps force required to extend the knee during an isokinetic extension cycle generating a constant extension moment after non-hinged and hinged total knee arthroplasty. METHODS: Human knee specimens were tested in a kinematic knee simulator under physiological conditions, after implantation of two types of non-hinged cruciate retaining prosthesis (Gemini; Link, Germany and Interax I.S.A.; Stryker, Ireland) and a hinged prosthesis (Rotations-Knie; Link, Germany). During simulation of an extension cycle from 120 degrees knee flexion to full extension, the change in quadriceps force to produce the constant extension moment of 31 Nm was dynamically measured using a load cell attached to the quadriceps tendon. RESULTS: After implantation of the non-hinged pros-theses, there was no alteration in maximum quadriceps force in knee flexion compared to physiological conditions, but alteration occurred at lower flexion angle (p=0.002) and increased up to 1,257 (SD 273) N (p=0.04) in knee extension. Following implantation of the hinged prosthesis, there was no alteration in quadriceps extension force in flexion but it decreased to 690 (SD 81) N (p=0.003) in extension. INTERPRETATION: Hinged knee prostheses restore the quadriceps lever arm in knee flexion and improve the lever arm in knee extension due to higher constraint and knee joint stability. This would offer a potential advantage for patients with weak quadriceps strength by making it easier to stabilize the knee in full extension during walking.