Einfluss von NO-Donoren und NO-Donor-Produgs auf die Permeabilität eines Modells der Blut-Hirn-Schranke aus primären Schweinehirnendothelzellen

Ziel der Promotionsarbeit: Im Rahmen des Projektes "Entdeckung und Entwicklung von neuartigen Wirkstoffen und Trägersystemen für die Pharmakotherapie der Alzheimer-Demenz" der pharmazeutischen Institute der Friedrich-Schiller-Universität Jena wurde ein Modell benötigt, dass die Blut-Hirn-Schranke simuliert, so dass daran neue Substanzen auf ihre Eigenschaften getestet werden können. Dieses Modell soll Aussagen zur Penetrationsfähigkeit von Substanzen ermöglichen. Des weiteren ist mit einem Modell aus Endothelzellen die Untersuchung von Einflüssen neuer Substanzen auf die Dichtigkeit der Monolayer möglich. An diesem Modell sollen die Einflüsse der unterschiedlichen NO-Donoren und NO-Donor-Prodrugs auf die Dichtigkeit der Endothelzellmonolayer untersucht werden. Durchführung: Gemessen wurde der Fluss von Carboxyfluorescein über einen Endothelzellmonolayer aus primären Schweinehirnendothelzellen. Dieser Monolayer wurde mit NO-Donoren, NO-Donor- Prodrugs oder einer Kombination aus NO-Donoren und Methylenblau oder NO-Donor- Prodrugs und Methylenblau behandelt und der dadurch beeinflusste Fluss von Carboxyfluorescein gemessen

Microscopic and Molecular Characterization of the Prehaustorial Resistance against Wheat Leaf Rust (Puccinia triticina) in Einkorn (Triticum monococcum)

Puccinia triticina f. sp. tritici (Eriks.), the causal agent of leaf rust, causes substantial yield losses in wheat production. In wheat many major leaf rust resistance genes have been overcome by virulent races. In contrast, the prehaustorial resistance (phr) against wheat leaf rust detected in the diploid wheat Einkorn (Triticum monoccocum var. monococcum) accession PI272560 confers race-independent resistance against isolates virulent on accessions harbouring resistance genes located on the A-genome of Triticum aestivum. Phr in PI272560 leads to abortion of fungal development during the formation of haustorial mother cells and to increased hydrogen peroxide concentration in comparison to the susceptible accession 36554 (Triticum boeoticum ssp. thaoudar var. reuteri). Increased peroxidase and endochitinase activity was detected in PI272560 within 6 hours after inoculation (hai). Comparative transcriptome profiling using Massive Analysis of cDNA Ends (MACE) in infected and non-infected leaves detected 14220 differentially expressed tags in PI272560 and 15472 in accession 36554. Of these 2908 and 3004, respectively, could be assigned to Gene Ontology (GO) categories of which 463 were detected in both accessions and 311 were differentially expressed between the accessions. In accordance with the concept of nonhost resistance in PI272560, genes with similarity to peroxidases, chitinases, β-1,3-glucanases and other pathogenesis-related genes were up-regulated within the first 8 hai, whereas up-regulation of such genes was delayed in 36554. Moreover, a Phosphoribulokinase gene contributing to nonhost resistance in rice against stripe rust was exclusively expressed in the resistant accession PI272560. Gene expression underpinned physiological and phenotypic observations at the site of infection and are in accordance with the concept of nonhost resistance

Characterization and Assessment of a Novel Plate and Frame MD Module for Single Pass Wastewater Concentration−FEED Gap Air Gap Membrane Distillation

Membrane distillation (MD) is an up and coming technology for concentration and separation on the verge of reaching commercialization. One of the remaining boundaries is the lack of available full-scale MD modules and systems suitable to meet the requirements of potential industrial applications. In this work a new type of feed gap air gap MD (FGAGMD) plate and frame module is introduced, designed and characterized with tap water and NaCl–H2O solution. The main feature of the new channel configuration is the separation of the heating and cooling channel from the feed channel, enabling a very high recovery ratio in a single pass. Key performance indicators (KPIs) such as flux, gained output ratio (GOR), recovery ratio and thermal efficiency are used to analyze the performance of the novel module concept within this work. A recovery rate of 93% was reached with tap water and between 32–53% with salt solutions ranging between 117 and 214 g NaCl/kg solution with this particular prototype module. Other than recovery ratio, the KPIs of the FGAGMD are similar to those of an air gap membrane distillation (AGMD) channel configuration. From the experimental results, furthermore, a new MD KPI was defined as the ratio of heating and cooling flow to feed flow. This RF ratio can be used for optimization of the module design and efficiency.BMBF, 02WAV1406E, WavE - Verbundprojekt HighCon: Konzentrate aus der Abwasserwiederverwendung, Teilprojekt

Nanoparticulate CpG Immunotherapy in RAO- Affected Horses: Phase I and IIa Study

Background: Recurrent airway obstruction (RAO), an asthma-like disease, is 1 of the most common allergic diseases in horses in the northern hemisphere. Hypersensitivity reactions to environmental antigens cause an allergic inflammatory response in the equine airways. Cytosine-phosphate-guanosine-oligodeoxynucleotides (CpG-ODN) are known to direct the immune system toward a Th1-pathway, and away from the pro-allergic Th2-line (Th2/Th1-shift). Gelatin nanoparticles (GNPs) are biocompatible and biodegradable immunological inert drug delivery systems that protect CpG-ODN against nuclease degeneration. Preliminary studies on the inhalation of GNP-bound CpG-ODN in RAO-affected horses have shown promising results. Objectives: The aim of this study was to evaluate the clinical and immunological effects of GNP-bound CpG-ODN in a double-blinded, placebo-controlled, prospective, randomized clinical trial and to verify a sustained effect post-treatment. Animals and Methods: Twenty-four RAO-affected horses received 1 inhalation every 2 days for 5 consecutive administrations. Horses were examined for clinical, endoscopic, cytological, and blood biochemical variables before the inhalation regimen (I), immediately afterwards (II), and 4 weeks post-treatment (III). Results: At time points I and II, administration of treatment rather than placebo corresponded to a statistically significant decrease in respiratory effort, nasal discharge, tracheal secretion, and viscosity, AaDO2 and neutrophil percentage, and an increase in arterial oxygen pressure. Conclusion and Clinical Importance: Administration of a GNP-bound CpG-ODN formulation caused a potent and persistent effect on allergic and inflammatory-induced clinical variables in RAO-affected horses. This treatment, therefore, provides an innovative, promising, and well-tolerated strategy beyond conventional symptomatic long-term therapy and could serve as a model for asthma treatment in humans

Immunomodulatory asthma therapy in the equine animal model: A dose‐response study and evaluation of a long‐term effect

Introduction Equine asthma represents a naturally occurring animal model for human allergic neutrophilic asthma. Inhalative nanoparticle‐bound cytosine‐phosphate‐guanosine (CpG‐GNP) immunotherapy, independent of specific allergens, has already shown promising clinical and immunological results in previous studies and offers the possibility to treat the underlying cause of the disease. This study analyses the relationship between dose and response, and evaluates a possible long‐term effect. Methods In the prospective, randomised, double‐blind clinical field study, 29 horses suffering from equine asthma received 10 inhalation treatments with either 187.5 µg CpG‐GNP (CpG single dose [CpGsd]; n = 11), 375 µg CpG‐GNP double dose (CpG double dose [CpGdd]; n = 9) (q48h for 20 days) or 1600 µg beclomethasone (n = 9) (q24h for 10 days). Each horse was examined three times: before the treatment (I), immediately after the 10 inhalations (II), and 8 weeks after the final inhalation (III). The three groups were compared according to clinical and laboratory parameters. The study examined the sustainability of the long‐term effect of the treatment after 8 weeks, as well as the tolerability of the formula as a double dose. Results The CpGsd resulted in a significant improvement in 82% of the parameters, the CpGdd in 72%. In the long‐term evaluation, the CpGsd showed a significant improvement in 100% of the parameters in comparison to the initial values, the CpGdd in 67%. On the immunological level, the bronchoalveolar lavage revealed a significant reduction of IL‐4, IL‐8, and interferon‐γ. Conclusion Both CpG groups displayed significant improvements in clinical and laboratory parameters, especially regarding the long‐term effect of CpGsd. Doubling the CpG dose did not result in any improvement in comparison to the original single dose. On the immunological level, an anti‐inflammatory, as well as an immunomodulatory effect, apart from a Th2‐dominated immune response, could be observed. This immunomodulatory inhalation treatment could indicate a new possibility for human allergic asthma therapy

A comparison of nanoparticullate CpG immunotherapy with and without allergens in spontaneously equine asthma-affected horses, an animal model

Introduction: New therapeutic strategies to modulate the immune response of human and equine allergic asthma are still under extensive investigation. Immunomodulating agents stimulating T-regulatory cells offer new treatment options beyond conventional symptomatic treatment or specific immunotherapy for human and equine allergic airway diseases, with the goal of a homoeostatic T-helper cell balance. The aim of this study was to evaluate the effects of a nebulized gelatin nanoparticle-CpG formulation (CpG-GNP) with and without specific allergens for the treatment of spontaneous allergic equine asthma as a model for human asthma. Methods: Twenty equine asthma-affected horses were treated either with CpG-GNP alone or CpG-GNP with allergens. Two specific allergens were selected for each horse based on history and an in-vitro test. Each horse received seven administrations of the respective nebulized composition and was examined before treatment, immediately after and 6 weeks after the treatment course. Results: Clinical parameters such as breathing rate, indirect interpleural measurement, arterial blood gases, amount of tracheal mucus and percentage of neutrophils and cytokines in tracheal washes and serum samples were evaluated. Treatment with CpG-GNP alone as well as in combinations with relevant allergens resulted in clinical improvement of nasal discharge, breathing rate, amount of secretion and viscosity, neutrophil percentage and partial oxygen pressure directly after and 6 weeks after treatment. There were no significant differences between the two treatments in clinical parameters or local cytokine profiles in the tracheal wash fluid (IL-10, IFN-g, and IL-17). IL-4 concentrations decreased significantly in both groups. Conclusion: Nonspecific CpG-GNP-based immunotherapy shows potential as a treatment for equine and possibly also human allergic asthma

Mechanisms contributing to visceral hypersensitivity : focus on splanchnic afferent nerve signalling

This is the peer reviewed version of the following article: Deiteren, A., De Man, J. G., Keating, C., Jiang, W., De Schepper, H. U., Pelckmans, P. A., Francque, S. M. and De Winter, B. Y. (2015), Mechanisms contributing to visceral hypersensitivity: focus on splanchnic afferent nerve signaling. Neurogastroenterology & Motility, 27: 1709–1720, which has been published in final form at doi:10.1111/nmo.12667. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.Visceral hypersensitivity is a main characteristic of functional bowel disorders and is mediated by both peripheral and central factors. We investigated whether enhanced splanchnic afferent signaling in vitro is associated with visceral hypersensitivity in vivo in an acute and postinflammatory rat model of colitis.Peer reviewedFinal Accepted Versio

Assessing incomplete neutralization of final devoicing in German

It has been claimed that the long established neutralization of the voicing distinction in domain final position in German is phonetically incomplete. However, many studies that have advanced this claim have subsequently been criticized on methodological grounds, calling incomplete neutralization into question. In three production experiments and one perception experiment we address these methodological criticisms. In the first production study, we address the role of orthography. In a large scale auditory task using pseudowords, we confirm that neutralization is indeed incomplete and suggest that previous null results may simply be due to lack of statistical power. In two follow-up production studies (Experiments 2 and 3), we rule out a potential confound of Experiment 1, namely that the effect might be due to accommodation to the presented auditory stimuli, by manipulating the duration of the preceding vowel. While the between-items design (Experiment 2) replicated the findings of Experiment 1, the between-subjects version (Experiment 3) failed to find a statistically significant incomplete neutralization effect, although we found numerical tendencies in the expected direction. Finally, in a perception study (Experiment 4), we demonstrate that the subphonemic differences between final voiceless and devoiced stops are audible, but only barely so. Even though the present findings provide evidence for the robustness of incomplete neutralization in German, the small effect sizes highlight the challenges of investigating this phenomenon. We argue that without necessarily postulating functional relevance, incomplete neutralization can be accounted for by recent models of lexical organization. (C) 2014 Elsevier Ltd. All rights reserved

Structural and dynamic characterization of Li12Si7 and Li12Ge7 using solid state NMR

Local environments and lithium ion dynamics in the binary lithium silicide Li12Si7, and the analogous germanium compound have been characterized by detailed 6Li, 7Li, and 29Si variable temperature static and magic-angle spinning (MAS) NMR experiments. In the MAS-NMR spectra, individual lithium sites are generally well-resolved at temperatures below 200 K, whereas at higher temperatures partial site averaging is observed on the kHz timescale. The observed lithium chemical shift ranges of up to 60 ppm indicate a significant amount of electronic charge stored on the lithium species, consistent with the expectation of the extended Zintl–Klemm–Bussmann concept used for the theoretical description of lithium silicides. Furthermore the strongly diamagnetic chemical shifts observed for the lithium ions situated directly above the five-membered Si5 rings suggest the possibility of aromatic ring currents in these structural elements. This assignment is confirmed further by 29Si{7Li} CPMAS-heteronuclear correlation experiments. The 29Si MAS-NMR spectra of Li12Si7, aided by 2-D J-resolved spectroscopy, are well suited for differentiating between the individual sites within the silicon framework, while further detailed connectivity information is available on the basis of 2-D INADEQUATE and radio frequency driven recoupling (RFDR) spectra. Variable temperature static 7Li NMR spectra reveal the onset of strong motional narrowing effects, illustrating high lithium ionic mobilities in both of these compounds.DFG (Ec168/9-1