Rosellinia (Xylariaceae) is a large, cosmopolitan genus comprising over 130 species that have been defined based mainly on the morphology of their sexual morphs. The genus comprises both lignicolous ...and saprotrophic species that are frequently isolated as endophytes from healthy host plants, and important plant pathogens. In order to evaluate the utility of molecular phylogeny and secondary metabolite profiling to achieve a better basis for their classification, a set of strains was selected for a multi-locus phylogeny inferred from a combination of the sequences of the internal transcribed spacer region (ITS), the large subunit (LSU) of the nuclear rDNA, beta-tubulin (TUB2) and the second largest subunit of the RNA polymerase II (RPB2). Concurrently, various strains were surveyed for production of secondary metabolites. Metabolite profiling relied on methods with high performance liquid chromatography with diode array and mass spectrometric detection (HPLC-DAD/MS) as well as preparative isolation of the major components after re-fermentation followed by structure elucidation using nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry (HR-MS). Two new and nine known isopimarane diterpenoids were identified during our mycochemical studies of two selected Dematophora strains and the metabolites were tested for biological activity. In addition, the nematicidal cyclodepsipeptide PF1022 A was purified and identified from a culture of Rosellinia corticium, which is the first time that this endophyte-derived drug precursor has been identified unambiguously from an ascospore-derived isolate of a Rosellinia species. While the results of this first HPLC profiling were largely inconclusive regarding the utility of secondary metabolites as genus-specific chemotaxonomic markers, the phylogeny clearly showed that species featuring a dematophora-like asexual morph were included in a well-defined clade, for which the genus Dematophora is resurrected. Dematophora now comprises all previously known important plant pathogens in the genus such as D. arcuata, D. bunodes, D. necatrix and D. pepo, while Rosellinia s. str. comprises those species that are known to have a geniculosporium-like or nodulisporium-like asexual morph, or where the asexual morph remains unknown. The extensive morphological studies of L.E. Petrini served as a basis to transfer several further species from Rosellinia to Dematophora, based on the morphology of their asexual morphs. However, most species of Rosellinia and allies still need to be recollected in fresh state, cultured, and studied for their morphology and their phylogenetic affinities before the infrageneric relationships can be clarified.
Although successful in the structural determination of ordered biomolecules, the spectroscopic investigation of oligopeptides in solution is hindered by their complex and rapidly changing ...conformational ensemble. The measured circular dichroism (CD) spectrum of an oligopeptide is an ensemble average over all microstates, severely limiting its interpretation, in contrast to ordered biomolecules. Spectral deconvolution methods to estimate the secondary structure contributions in the ensemble are still mostly based on databases of larger ordered proteins. Here, we establish how the interpretation of CD spectra of oligopeptides can be enhanced by the ability to compute the same observable from a set of atomic coordinates. Focusing on two representative oligopeptides featuring a known propensity toward an α-helical and β-hairpin motif, respectively, we compare and cross-validate the structural information coming from deconvolution of the experimental CD spectra, sequence-based de novo structure prediction, and molecular dynamics simulations based on enhanced sampling methods. We find that small conformational variations can give rise to significant changes in the CD signals. While for the simpler conformational landscape of the α-helical peptide de novo structure prediction can already give reasonable agreement with the experiment, an extended ensemble of conformers needs to be considered for the β-hairpin sequence.
Rosellinia (Xylariaceae) is a large, cosmopolitan genus comprising over 130 species that have been defined based mainly on the morphology of their sexual morphs. The genus comprises both lignicolous ...and saprotrophic species that are frequently isolated as endophytes from
healthy host plants, and important plant pathogens. In order to evaluate the utility of molecular phylogeny and secondary metabolite profiling to achieve a better basis for their classification, a set of strains was selected for a multi-locus phylogeny inferred from a combination of the sequences
of the internal transcribed spacer region (ITS), the large subunit (LSU) of the nuclear rDNA, beta-tubulin (TUB2) and the second largest subunit of the RNA polymerase II (RPB2). Concurrently, various strains were surveyed for production of secondary metabolites. Metabolite profiling
relied on methods with high performance liquid chromatography with diode array and mass spectrometric detection (HPLC-DAD/MS) as well as preparative isolation of the major components after re-fermentation followed by structure elucidation using nuclear magnetic resonance (NMR) spectroscopy
and high resolution mass spectrometry (HR-MS). Two new and nine known isopimarane diterpenoids were identified during our mycochemical studies of two selected Dematophora strains and the metabolites were tested for biological activity. In addition, the nematicidal cyclodepsipeptide
PF1022 A was purified and identified from a culture of Rosellinia corticium, which is the first time that this endophyte-derived drug precursor has been identified unambiguously from an ascospore-derived isolate of a Rosellinia species. While the results of this first HPLC profiling
were largely inconclusive regarding the utility of secondary metabolites as genus-specific chemotaxonomic markers, the phylogeny clearly showed that species featuring a dematophora-like asexual morph were included in a well-defined clade, for which the genus Dematophora is resurrected.
Dematophora now comprises all previously known important plant pathogens in the genus such as D. arcuata, D. bunodes, D. necatrix and D. pepo, while Rosellinia s. str. comprises those species that are known to have a geniculosporium-like or nodulisporium-like
asexual morph, or where the asexual morph remains unknown. The extensive morphological studies of L.E. Petrini served as a basis to transfer several further species from Rosellinia to Dematophora, based on the morphology of their asexual morphs. However, most species of Rosellinia
and allies still need to be recollected in fresh state, cultured, and studied for their morphology and their phylogenetic affinities before the infrageneric relationships can be clarified.
The purpose of the study is to describe some rare and hitherto unreported uni- and bilateral anomalies of the orbital rectus muscles (duplication, triplication, accessory bellies, interrectal ...muscular bridges, false insertion) which were found by chance during the dissections of three cadavers (one adult, two fetuses). The levator palpebrae superioris muscles in the same specimens exhibited some variations (medial and lateral muscular slips) and anomalies (bipartite and unipartite levator). Cross-sections of interrectal bridges in the orbit of an adult were investigated histologically. Numerous nerves were distributed in the interstitium; groups of lipofuscin granules were found in all the myocytes. An attempt is made to explain these anomalies and variations through consideration of developmental and comparative anatomy. The relevance of these anomalies in coronal sections such as those acquired from CT and MRI is discussed.
The purpose of the study is to describe some rare and hitherto unreported uni- and bilateral anomalies of the orbital rectus muscles (duplication, triplication, accessory bellies, interrectal ...muscular bridges, false insertion) which were found by chance during the dissections of three cadavers (one adult, two fetuses). The levator palpebrae superioris muscles in the same specimens exhibited some variations (medial and lateral muscular slips) and anomalies (bipartite and unipartite levator). Cross-sections of interrectal bridges in the orbit of an adult were investigated histologically. Numerous nerves were distributed in the interstitium; groups of lipofuscin granules were found in all the myocytes. An attempt is made to explain these anomalies and variations through consideration of developmental and comparative anatomy. The relevance of these anomalies in coronal sections such as those acquired from CT and MRI is discussed.
Abstract
The European maritime transport policy recognizes the importance of the waterborne transport systems as key elements for sustainable growth in Europe. A major goal is to transfer more than ...50% of road transport to rail or waterways within 2050. To meet this challenge waterway transport needs to get more attractive and overcome its disadvantages. Therefore, it is necessary to develop new knowledge and technology and find a completely new approach to short sea and inland waterways shipping. A key element in this is automation of ships, ports and administrative tasks aligned to requirements of different European regions. One main goal in the AEGIS project is to increase the efficiency of the waterways transport with the use of higher degrees of automation corresponding with new and smaller ship types to reduce costs and secure higher frequency by feeders and provide multimodal green logistics solutions combining short sea shipping with rail and road transport.
Recent clinical reports suggest that the application of an autologous blood plasma enriched with thrombocytes by centrifugal concentration (platelet-rich plasma: PRP) can enhance the formation of new ...bone. There are very fewin vitro or in vivo studies published on the efficiency of PRP. In this project a three dimensional cell culture system was used to compare PRP and rhBMP-2 in vitro. Marrow derived bone forming cells from Spraque–Dawley (SD) rats were seeded on porous collagenous carriers (d=5mm, h=3mm) at a density of 4×104 cells/carrier and exposed to different concentrations of PRP (platelet counts from 2.5×108–1.6×107 platelets/culture), rhBMP-2 (300ng) or plasma poor in thrombocytes (platelet-poor plasma, PPP). Cultures without additional supplements were used as controls. During a culture period of 21 days cell proliferation, alkaline phosphatase activity (ALP) and calcium content (days 18, 21) were measured in 3 day intervals.
PRP showed a dose dependent stimulation of cell proliferation, while reducing ALP activity and calcium deposition in the culture. BMP-2 led to an opposite cell response and induced the highest ALP activity and mineral deposition. These data suggest that PRP inhibited osteogenic differentiation of marrow derived pre-osteoblasts in a dose dependent manner. PRP is not a substitute for BMP-2 in osteogenic induction.
In strong-field ionization processes, two-color laser fields are frequently used for controlling sub-cycle electron dynamics via the relative phase of the laser fields. Here we apply this technique ...to velocity map imaging spectroscopy using an unconventional orientation with the polarization of the ionizing laser field perpendicular to the detector surface and the steering field parallel to it. This geometry allows not only to image the phase-dependent photoelectron momentum distribution (PMD) of low-energy electrons that interact only weakly with the ion (direct electrons), but also to investigate the low yield of higher-energy rescattered electrons. Phase-dependent measurements of the PMD of neon and xenon demonstrate control over direct and rescattered electrons. The results are compared with semi-classical calculations in three dimensions including elastic scattering at different orders of return and with solutions of the three-dimensional time-dependent Schrödinger equation.
Atmospheric aerosol particles are activated and grow into drops during the formation of a cloud. Subsequently, they are delivered from the dissipating cloud drops back to the atmosphere. During the ...cloud lifetime, the drops scavenge water‐soluble trace gases, leading to an increase in size and solubility of the particles emerging from the evaporating cloud drops. This processing of aerosol particles by clouds has an influence on the microphysics of the following cloud and its probability to rain as well as on the cooling effect of the direct and indirect aerosol forcing of climate. To measure the cycling history (particle activation and gas scavenging in drops followed by processing of the activated particles followed by emerging processed particles) of continental aerosol particles passing through cloud drops of different sizes, a new method is developed and applied during three field experiments carried out on the Mount Kleiner Feldberg/Ts., Germany, in 1990, 1993, and 1995. The typical droplet spectra of most of the observed stratus clouds is weakly bimodal, with mode 1 at drop sizes between 3 and 5 μm and mode 2 between 5 and 10 μm radius. Cloud drops in this overall size range are subject to growth only by condensation, while coalescence can be neglected. Therefore the observed processing is related solely to gas scavenging and in‐cloud chemical reactions. We found that the processing of particles is different for the two modes of the cloud drop size spectrum: Small activated particles mostly grow to the small drops of mode 1, while larger particles can grow further to the larger drops of mode 2. Likewise, the mass of scavenged gas is, on average, lower for the small than for the larger drops. Vice versa, the ratio of scavenged gas to particle mass, the parameter quantifying the particle processing, is, on average, found to be higher in the small drop mode containing the smaller particles. The reason is that the degree of processing is mainly inversely linked to the mass of the activated particles. Therefore the strongest modification of particles takes place in smaller drops and affects mainly the smaller activated particles (r ap ≤ 0.1 μm). Their radii can increase by up to a factor of 3 and, consequently, their nucleation as well as radiative properties change significantly. The consequence for the aerosol climate forcing is that the cooling can be, to an unknown extent, intensified with increasing atmospheric amount of water‐soluble trace gases such as HNO3, NH3, and SO2, counteracting the warming effect of the greenhouse gases.
Durch gentechnische Modifikationen der N-terminalen Aminosäuresequenz von BMP-2 lässt sich die Heparinbindung und damit die lokale Verweildauer am Implantationsort gezielt beeinflussen. Zwei Mutanten ...mit verstärkter (T3, T4) und eine Mutante mit nahezu aufgehobener Heparinbindung (EHBMP-2) sollten auf ihre Osteoinduktivität in vivo überprüft und mit dem Wildtyp von BMP-2 verglichen werden. Unterschiedliche Konzentrationen (0,25-4 µg) der Proteine wurden auf zylindrische Kollagenträger (Durchmesser 5 mm, Höhe 10 mm) aufgebracht und in die Oberschenkelmuskulatur implantiert. Die Knochenbildung wurde mittels radiologischer Verlaufskontrollen quantifiziert und nach 28 Tagen histologisch charakterisiert. Durch die Aufhebung der Heparinbindung wird eine deutliche Reduktion der Osteoinduktivität in vivo bewirkt. Die Verstärkung der Heparinbindung führt dagegen zu einer Steigerung der Knocheninduktion, die beobachtete Knochenbildung findet schneller und in größerem Ausmaß statt. Durch gentechnische Modifikationen ist es gelungen, erstmalig BMP-Mutanten herzustellen, die eine verstärkte Bindung an Komponenten der extrazellulären Matrix (Heparin) aufweisen. Durch die hierdurch verlängerte Verweildauer am Implantationsort besitzen diese Mutanten eine gegenüber dem Wildtyp gesteigerte Osteoinduktivität. The alteration of the N-terminal amino acid sequence of BMP-2 allows modification of heparin binding of the new protein. This leads to a change in the local retention time at the site of implantation. Mutants with increased (T3, T4) and with no binding (EHBMP-2) to heparin were assessed for their osteoinductivity in vivo and compared with the wild type BMP-2. Cylindrical collagenous carriers (diameter = 5 mm, height = 10) were loaded with different concentrations (0.25-4 µg) of the proteins. Following intramuscular implantation into the hind legs, the bone formation was measured in radiographic follow-ups. After 28 days the newly formed bone was characterized histologically. Elimination of the heparin binding leads to massive reduction in osteoinductivity. On the other hand, an increase in the heparin binding leads to enhancement in the osteoinductive properties, resulting in faster bone formation with a higher yield. It could be shown for the first time that modifications of BMP-2 by gene technology can lead to proteins with enhanced binding to components of the extracellular matrix. The resulting prolonged retention time at the implantation site results in an increased osteoinductivity compared with the wild type.PUBLICATION ABSTRACT
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