Fecal specimens from 130 different mares were collected from an endemic farm for 2 consecutive years at 4 different times pre- and post-foaling (41 mares contributed data in both years). A modified NANAT agar medium was used to quantitatively culture 1-g aliquots of the mare feces without inhibition of growth of Rhodococcus equi. Once the R. equi in the mare feces were quantified and the total concentrations of R. equi determined, a colony immunoblot procedure was performed to detect the presence of the virulence-associated protein antigen on the isolates. This allowed for the proportion and concentration of virulent R. equi to be determined. Foals that were found to have ultrasonographic evidence of peripheral pulmonary abscessation or consolidation underwent aseptic trans-cutaneous tracheobronchial aspiration. Positive results of TBA were used to categorize foals as affected with R. equi pneumonia. R. equi pneumonia developed in 31% of the foals. Shedding of virulent R. equi was observed in at least 1 sampling period for every mare examined, and >33% were culture-positive during all sampling periods. However, significant differences were not observed in either the fecal concentrations of total or virulent R. equi from dams of affected foals compared to dams of unaffected foals. No significant temporal changes in the fecal concentrations of R. equi were observed. It was concluded that dams of affected foals do not shed more R. equi in feces than do dams of unaffected foals, indicating that heavier shedding by particular mares does not explain infection in their foals. However, the finding that virulent R. equi were excreted in the feces of all sampled mares indicates that mares are likely an important source of R. equi for their surrounding environment.

Fecal specimens from 130 different mares were collected from an endemic farm for 2 consecutive years at 4 different times pre- and post-foaling (41 mares contributed data in both years). A modified NANAT agar medium was used to quantitatively culture 1-g aliquots of the mare feces without inhibition of growth of Rhodococcus equi. Once the R. equi in the mare feces were quantified and the total concentrations of R. equi determined, a colony immunoblot procedure was performed to detect the presence of the virulence-associated protein antigen on the isolates. This allowed for the proportion and concentration of virulent R. equi to be determined. Foals that were found to have ultrasonographic evidence of peripheral pulmonary abscessation or consolidation underwent aseptic trans-cutaneous tracheobronchial aspiration. Positive results of TBA were used to categorize foals as affected with R. equi pneumonia. R. equi pneumonia developed in 31% of the foals. Shedding of virulent R. equi was observed in at least 1 sampling period for every mare examined, and >33% were culture-positive during all sampling periods. However, significant differences were not observed in either the fecal concentrations of total or virulent R. equi from dams of affected foals compared to dams of unaffected foals. No significant temporal changes in the fecal concentrations of R. equi were observed. It was concluded that dams of affected foals do not shed more R. equi in feces than do dams of unaffected foals, indicating that heavier shedding by particular mares does not explain infection in their foals. However, the finding that virulent R. equi were excreted in the feces of all sampled mares indicates that mares are likely an important source of R. equi for their surrounding environment.

Zugl.: Hannover, Tierärztliche Hochsch., Diss., 2007

Connexin32- defiziente Mäuse stellen ein Mausmodell für eine Form der hereditären peripheren Neuropahtie dar. Es konnte gezeigt werden, dass Makrophagen, möglicherweise aktiviert durch MCP-1, die Demyelinisierung in Connexin32-defizienten Mäusen vermitteln. Diese Arbeit untersucht mögliche Signaltransduktionswege, die in den peripheren Nerven Connexin32- defizienter Mäuse aktiviert sein könnten und damit in Zusammenhang mit der Genexpression von MCP-1 und/oder Makrophagen-Aktivierung stehen könnten. / Connexin32 deficient mice are a well established mouse model for the hereditary neuropathy known as Charcot-Marie-Tooth disease. Recently it was shown that macrophages function as the mediators of demyelination in peripheral nerves. Additionally there was found an icreased gene expression of MCP-1 in peripheral nerves of these animals. This study tries to identify un activated signal transduction pathway in peripheral nerves of Connexin32 deficient mice that may lead to the increased gene expression of MCP-1 and increased number of macrophages in peripheral nerves of Connexin32 deficient mice.

La prévalence de l’allergie alimentaire est en constante augmentation. Parallèlement les consommations alimentaires et les aliments eux même évoluent. L’aliment, de plus en plus soumis aux traitements industriels, peut présenter une allergénicité modifiée. L’objet de cette thèse sera d’évaluer l’influence des différents procédés industriels sur l’immunoréactivité des protéines alimentaires modifiées. Les différentes études présentées dans l’ouvrage aborderont l’effet de la texturation sur les protéines de soja ou de l’extrusion sur les protéines de lin (traitement thermique), l’effet de l’incorporation de nouveaux ingrédients dans l’alimentation moderne (inuline, lin). L’allergénicité des protéines recombinantes (process biotechnologique) sera abordée au travers de deux études : l’une proposera les protéines recombinantes de l’arachide comme outil diagnostique, en particulier pour le test d’activation des basophiles ; L’autre cherchera à démontrer l’innocuité allergénique d’un lysozymes recombinant humain, qui sera utilisé en alimentation humaine. L’étude de ces propriétés allergéniques sera réalisée par les tests in vitro (prick tests et test de provocation orale) et par les tests in vitro (IgEs). Des études par immuno-empreinte (immunoblot et dot blot) et par spectrométrie infrarouge permettront d’identifier les protéines modifiées. Enfin des tests cellulaires (test d’activation des basophiles) seront également réalisés. / The prevalence of the food allergy is in constant increase. At the same time food consumptions and food they even evolve. The food, more and more subjected to the industrial treatments, can present a modified allergenicity. The aim of this thesis will be to estimate the influence of the various industrial processes on the immunoreactivity of modified food proteins. The various studies presented in the work will approach the effect of the texturisation on proteins of soy or the extrusion on flaxseed proteins (heat treatment), the effect of the incorporation of new ingredients in the modern food (inuline, flaxseed). The allergenicity of recombinant proteins (biotechnological process) will be approached through two studies: the one will propose peanut recombinant proteins as a diagnostic tool, in particular for the basophil activation test (BAT); other one will try to demonstrate the allergenic harmlessness of a recombinant human lysozyme, which will be used in human food. The study of these allergenic properties will be realized by the in vivo tests (prick tests) and by the in vitro tests (IgEs). Studies by immunoblot and by infrared spectrometry will allow identifying modified proteins. Finally cellular tests (BAT) will be also performed.

Die Spinale Muskelatrophie (SMA) ist mit einer Inzidenz von 1:6000 die zweithäufigste autosomal-rezessive Erbkrankheit im frühen Kindesalter. Die durch den Verlust des SMN- (survival of motoneuron) Gens reduzierte SMN Protein Expression führt zu einer Degeneration der spinalen Motoneurone mit proximal betonter Muskelschwäche. Deshalb zielen erste Therapieversuche darauf ab, diese zu erhöhen. Es war gezeigt worden, dass durch den Einsatz von Histon Deacetylase Inhibitoren (HDAC) in neuronalen Kontroll Zellen und in nicht neuronalen Zellen von SMA Patienten die SMN Protein Expression signifikant gesteigert werden konnte. In der vorgelegten Arbeit wurde untersucht, ob die HDAC Inhibitoren Valproat, SAHA und FK228 Einfluss auf die SMN Protein Expression in kortikalen neuronalen Vorläuferzellen (NSC), auf primär embryonale Fibroblasten (PMEF) und auf die morphologischen Veränderungen in primär kultivierten embryonalen Motoneuronen eines Mausmodells der SMA haben. Es konnte eine signifikante Steigerung der SMN Protein Expression durch den Einsatz von Valproat und FK228 in kortikalen neuronalen Vorläuferzellen nachgewiesen werden. Es ergab sich jedoch kein Einfluss auf die SMN Protein Expression in primär embryonalen Fibroblasten. Bei NSCs und primär kultivierten embryonalen Motoneuronen wirkten die HDAC Inhibitoren Valproat und FK228 konzentrationsabhängig toxisch auf das Überleben, die Länge der Axone und die Größe der Wachstumskegel. Es konnte kein positiver Einfluss auf die morphologischen Veränderungen des Mausmodells gesehen werden. Zusammenfassend zeigte sich in der vorgelegten Arbeit ein positiver Effekt auf die SMN Protein Expression durch den Einsatz von HDAC Inhibitoren, der jedoch mit einem toxischen Effekt auf die behandelten neuronalen Zellen einherging. / Spinal muscular atrophy (SMA) has an incidence of 1:6000 and is the second most common autosomal recessive hereditary disease in early childhood. The disease is characterized by the degeneration of spinal motor neurons with weakness of the proximal limb. This is caused by the deficiency of the SMN (survival of motor neuron) protein. Therefore therapeutical strategies aim to increase the SMN protein level. It has been shown that histone deacetylase inhibitors could increase SMN protein level in neuronal control cells and non-neuronal cells of SMA patients. The aim of the presented study was to investigate whether the HDAC inhibitors valproic acid, SAHA and FK228 had an effect on the SMN protein level in cortical neural progenitor cells or primary embryonic fibroblasts of a SMA mice model. The second question was if morphological pathologies in primary cultured embryonic motor neurons of this SMA mouse model could be altered. There was a significant increase in SMN protein level by the use of valproic acid and FK228 in cortical neuronal precursor cells. However, there was no effect on the SMN protein level in primary embryonic fibroblasts. In cortical neuronal precursor cells and primary cultured embryonic motor neurons, the HDAC inhibitors valproic acid and FK228 displayed and concentration-dependent toxic effect on the survival, axonal length and the size of the growth cone. No obvious influence on the morphological changes in the mice model could be seen. In conclusion a positive effect on the SMN protein level by the use of HDAC inhibitors could be detected, but with a toxic effect on neuronal cells at the same time.

Konstanz, Univ., Bachelorarb., 2007.

Techn. Universiẗat, Diss., 2003--Berlin.

Tierärztl. Hochsch., Diss., 2004--Hannover.

Bereits in Vorstudien konnte dargelegt werden, dass eine signifikante Korrelation zwischen der T-Zell-Zytokin-Antwort und der berufs- bzw. umweltbedingten Schimmelpilzbelastung besteht. Ziel der vorliegenden Studie war, eine mögliche Kombination von Biomarkern ausfindig zu machen, die veränderte T-Zell-Antworten auf A. fumigatus- Antigene bei beruflich Exponierten im Vergleich zu Kontrollprobanden/-innen vorhersagen kann. Um geeignete Marker für das Bio-Monitoring zu finden, wurden zur T-Zell-Aktivierung ein myzeliales A. fumigatus - Lysat und 12 proteinogene Antigene in ELISpot-Versuchen für die Signaturzytokine IFN-γ (TH1), IL-5 (TH2) und IL-17A (TH17) der Haupt-TH-Subpopulationen getestet. Es zeigten sich bei den Biolandwirten/-innen erwartungsgemäß erhöhte TH1- und TH2-Antworten auf die Mehrzahl der verwendeten spezifischen A. fumigatus-Antigene, die möglicherweise eine Schimmelpilzbelastung serologisch nachweisbar machen. Insbesondere die spezifischen A. fumigatus-Antigene Aspf22, CatB und CipC konnten eine Trennschärfe zwischen den beiden Kohorten hinsichtlich ihrer IFN-γ- und IL-5-Zytokinantwort erzielen. Unterschiede in der TH17-Antwort aufgrund chronischer beruflicher Sporenbelastung ohne Krankheitskorrelat konnten nicht explizit festgestellt werden. Weiterhin ergab sich, dass erhöhte TH2-Immunreaktionen, sofern sie mit einer adäquaten TH1-gerichteten Immunantwort einhergehen und damit eine ausgeglichene TH2/TH1-Balance besteht, nicht zwangsläufig zu Hypersensitivitätserkrankungen führen. Im Vergleich zu Langzeitexponierten wurden teilweise überlappende TH-Zellfrequenzen bei beruflich exponierten Biolandwirten/-innen ermittelt. Welche entscheidende Rolle Treg-Zellen bei der Eindämmung überschießender Immunantworten einnehmen, kann hieraus erahnt werden. / Preliminary studies have already set out a significant correlation between T-cell-cytokine-responses and professional and/or environmental mold contaminations. This study aimed for a possible combination of biomarkers to predict altered T-cell-responses to A. fumigatus- antigens in a defined professionally exposed cohort in comparison with controls. To find out suitable markers for the biomonitoring, T-cell-activation was performed with a mycelial A. fumigatus- lysate and 12 proteinaceous antigens in ELISpot assays with look at the main signature cytokines of T-helper cell subsets IFN-γ (TH1), IL-5 (TH2) and IL-17A (TH17). As immunoglobulins play an important role in pathophysiology and diagnostic of allergies, sera of the subjects were examined regarding IgE-specific antibodies against mycelial A. fumigatus - lysate and 12 proteinaceous antigens in Western Blot-technique. As expected, increased levels of TH1- and TH2-cytokine responses were found for a majority of examined specific A. fumigatus-antigens, which may help to verify mold exposure in a serological way. Differences in TH17-immune response in agricultural workers without a hypersensitivity disease has not been found. Furthermore, increased TH2-immune responses don’t necessarily lead to a pathologic value since it is followed by an adequate TH1-answer for upholding the TH2/TH1-balance. TH-frequencies of not occupational exposed individuals partly show overlaps with those of agricultural workers. Therefore, the important role of Treg-cells in containing extensive immune responses is conceivable. The specific A. fumigatus-antigens Aspf22, CatB and CipC are (in contrast to Aspergillus-lysate) able to drag selectivity in-between the two cohorts regarding their IFN- - and IL-5-cytokine response. Even though further investigations must be made, specific antigens are more suitable for exposition-analysis and, eventually, for the depiction of associated hypersensitivity diseases. The discrimination based on B-cell-triggered IgE- immune responses remains difficult, conceivably by reason of the subclinical status of the subjects. Solely Aspf3, which is already established in the Immuno-CAP-analysis, shows differential power.