Cloning and functional expression of the murine homologue of proteinase 3: implications for the design of murine models of vasculitis

AbstractAnti-neutrophil cytoplasmic autoantibodies recognizing conformational epitopes (c-ANCA) of proteinase 3 (PR3) from azurophil granules are a diagnostic hallmark in Wegener's granulomatosis (WG). Because a functional PR3 homologue has not been identified in rodents, it is difficult to assess immunopathological responses in rats or mice immunized with patients' derived c-ANCA or human PR3. Here we report the full length cDNA cloning and functional expression of murine PR3 in HMC-1 cells. Recombinant murine PR3 shows highly similar substrate specificities towards synthetic peptides and is inhibited by human α1-proteinase inhibitor like human PR3. However, neither human c-ANCA, rabbit sera nor mouse monoclonal antibodies to human PR3 recognize the murine homologue. Consequently, it is unlikely that disease observed in mice after immunization with c-ANCA or human PR3 is caused by pathogenic antibodies directed against mouse PR3. Recombinant human-mouse chimaeric variants will be a valuable new tool to localize the disease-specific immunodominant epitopes in human PR3

Quantitative DNA Analyses for Airborne Birch Pollen

Birch trees produce large amounts of highly allergenic pollen grains that are distributed by wind and impact human health by causing seasonal hay fever, pollen-related asthma, and other allergic diseases. Traditionally, pollen forecasts are based on conventional microscopic counting techniques that are labor-intensive and limited in the reliable identification of species. Molecular biological techniques provide an alternative approach that is less labor-intensive and enables identification of any species by its genetic fingerprint. A particularly promising method is quantitative Real-Time polymerase chain reaction (qPCR), which can be used to determine the number of DNA copies and thus pollen grains in air filter samples. During the birch pollination season in 2010 in Mainz, Germany, we collected air filter samples of fine (<3 μm) and coarse air particulate matter. These were analyzed by qPCR using two different primer pairs: one for a single-copy gene (BP8) and the other for a multi-copy gene (ITS). The BP8 gene was better suitable for reliable qPCR results, and the qPCR results obtained for coarse particulate matter were well correlated with the birch pollen forecasting results of the regional air quality model COSMO-ART. As expected due to the size of birch pollen grains (~23 μm), the concentration of DNA in fine particulate matter was lower than in the coarse particle fraction. For the ITS region the factor was 64, while for the single-copy gene BP8 only 51. The possible presence of so-called sub-pollen particles in the fine particle fraction is, however, interesting even in low concentrations. These particles are known to be highly allergenic, reach deep into airways and cause often severe health problems. In conclusion, the results of this exploratory study open up the possibility of predicting and quantifying the pollen concentration in the atmosphere more precisely in the future

Nanosized Multifunctional Polyplexes for Receptor-Mediated SiRNA Delivery

Although our understanding of RNAi and our knowledge on designing and synthesizing active and safe siRNAs significantly increased during the past decade, targeted delivery remains the major limitation in the development of siRNA therapeutics. On one hand, practical considerations dictate robust chemistry reproducibly providing precise carrier molecules. On the other hand, the multistep delivery process requires dynamic multifunctional carriers of substantial complexity. We present a monodisperse and multifunctional carrier system, synthesized by solid phase supported chemistry, for siRNA delivery in vitro and in vivo. The sequence-defined assembly includes a precise cationic (oligoethanamino)amide core, terminated at the ends by two cysteines for bioreversible polyplex stabilization, at a defined central position attached to a monodisperse polyethylene glycol chain coupled to a terminal folic acid as cell targeting ligand. Complexation with an endosomolytic influenza peptide-siRNA conjugate results in nanosized functional polyplexes of 6 nm hydrodynamic diameter. The necessity of each functional substructure of the carrier system for a specific and efficient gene silencing was confirmed. The nanosized polyplexes showed stability in vivo, receptor-specific cell targeting, and silencing of the EG5 gene in receptor-positive tumors. The nanosized appearance of these particles can be precisely controlled by the oligomer design (from 5.8 to 8.8 nm diameter). A complete surface charge shielding together with the high stability result in good tolerability in vivo and the absence of accumulation in nontargeted tissues such as liver, lung, or spleen. Due to their small size, siRNA polyplexes are efficiently cleared by the kidney

Visualization of interindividual differences in spinal dynamics in the presence of intraindividual variabilities

Surface topography systems enable the capture of spinal dynamic movement. A visualization of possible unique movement patterns appears to be difficult due to large intraclass and small inter-class variabilities. Therefore, we investigated a visualization approach using Siamese neural networks (SNN) and checked, if the identification of individuals is possible based on dynamic spinal data. The presented visualization approach seems promising in visualizing subjects in the presence of intraindividual variability between different gait cycles as well as day-to-day variability. Overall, the results indicate a possible existence of a personal spinal ‘fingerprint’. The work forms the basis for an objective comparison of subjects and the transfer of the method to clinical use cases

Analysis of the solar cycle and core rotation using 15 years of Mark-I observations:1984-1999. I. The solar cycle

High quality observations of the low-degree acoustic modes (p-modes) exist for almost two complete solar cycles using the solar spectrophotometer Mark-I, located at the Observatorio del Teide (Tenerife, Spain) and operating now as part of the Birmingham Solar Oscillations Network (BiSON). We have performed a Fourier analysis of 30 calibrated time-series of one year duration covering a total period of 15 years between 1984 and 1999. Applying different techniques to the resulting power spectra, we study the signature of the solar activity changes on the low-degree p-modes. We show that the variation of the central frequencies and the total velocity power (TVP) changes. A new method of simultaneous fit is developed and a special effort has been made to study the frequency-dependence of the frequency shift. The results confirm a variation of the central frequencies of acoustic modes of about 450 nHz, peak-to-peak, on average for low degree modes between 2.5 and 3.7 mHz. The TVP is anti-correlated with the common activity indices with a decrease of about 20% between the minimum and the maximum of solar cycle 22. The results are compared with those obtained for intermediate degrees, using the LOWL data. The frequency shift is found to increase with the degree with a weak l-dependence similar to that of the inverse mode mass. This verifies earlier suggestions that near surface effects are predominant.Comment: Accepted by A&A October 3 200

The non-detection of oscillations in Procyon by MOST: is it really a surprise?

We argue that the non-detection of oscillations in Procyon by the MOST satellite reported by Matthews et al. (2004) is fully consistent with published ground-based velocity observations of this star. We also examine the claims that the MOST observations represent the best photometric precision so far reported in the literature by about an order of magnitude and are the most sensitive data set for asteroseismology available for any star other than the Sun. These statements are not correct, with the most notable exceptions being observations of oscillations in alpha Cen A that are far superior. We further disagree that the hump of excess power seen repeatedly from velocity observations of Procyon can be explained as an artefact caused by gaps in the data. The MOST observations failed to reveal oscillations clearly because their noise level is too high, possibly from scattered Earthlight in the instrument. We did find an excess of strong peaks in the MOST amplitude spectrum that is inconsistent with a simple noise source such as granulation, and may perhaps indicate oscillations at roughly the expected level.Comment: 6 pages, accepted for publication in A&A Letter

a comparative experimental and numerical study of baroclinic wave dynamics

The differentially heated rotating annulus is a widely studied tabletop-size laboratory model of the general mid-latitude atmospheric circulation. The two most relevant factors of cyclogenesis, namely rotation and meridional temperature gradient are quite well captured in this simple arrangement. The radial temperature difference in the cylindrical tank and its rotation rate can be set so that the isothermal surfaces in the bulk tilt, leading to the formation of baroclinic waves. The signatures of these waves at the free water surface have been analyzed via infrared thermography in a wide range of rotation rates (keeping the radial temperature difference constant) and under different initial conditions. In parallel to the laboratory experiments, five groups of the MetStröm collaboration have conducted numerical simulations in the same parameter regime using different approaches and solvers, and applying different initial conditions and perturbations. The experimentally and numerically obtained baroclinic wave patterns have been evaluated and compared in terms of their dominant wave modes, spatio-temporal variance properties and drift rates. Thus certain “benchmarks” have been created that can later be used as test cases for atmospheric numerical model validation

Water uptake of subpollen aerosol particles: Hygroscopic growth, cloud condensation nuclei activation, and liquid-liquid phase separation

Pollen grains emitted from vegetation can release subpollen particles (SPPs) that contribute to the fine fraction of atmospheric aerosols and may act as cloud condensation nuclei (CCN), ice nuclei (IN), or aeroallergens. Here, we investigate and characterize the hygroscopic growth and CCN activation of birch, pine, and rapeseed SPPs. A high-humidity tandem differential mobility analyzer (HHTDMA) was used to measure particle restructuring and water uptake over a wide range of relative humidity (RH) from 2 % to 99.5 %, and a continuous flow CCN counter was used for size-resolved measurements of CCN activation at supersaturations (S) in the range of 0.2 % to 1.2 %. For both subsaturated and supersaturated conditions, effective hygroscopicity parameters, κ, were obtained by Köhler model calculations. Gravimetric and chemical analyses, electron microscopy, and dynamic light scattering measurements were performed to characterize further properties of SPPs from aqueous pollen extracts such as chemical composition (starch, proteins, DNA, and inorganic ions) and the hydrodynamic size distribution of water-insoluble material. All investigated SPP samples exhibited a sharp increase of water uptake and κ above ∼95 % RH, suggesting a liquid–liquid phase separation (LLPS). The HHTDMA measurements at RH >95 % enable closure between the CCN activation at water vapor supersaturation and hygroscopic growth at subsaturated conditions, which is often not achieved when hygroscopicity tandem differential mobility analyzer (HTDMA) measurements are performed at lower RH where the water uptake and effective hygroscopicity may be limited by the effects of LLPS. Such effects may be important not only for closure between hygroscopic growth and CCN activation but also for the chemical reactivity, allergenic potential, and related health effects of SPPs