<p>The blood-brain barrier restricts the access of many compounds, including therapeutic agents, to the brain. Several human studies indicate that nasal administration of hydrophilic compounds, such as peptides, can bypass the blood-brain barrier. The aims of this thesis were to develop and refine models for this direct nose-to-brain transfer.</p><p>In a mouse model, [³H]-dopamine was given as a unilateral nasal dose. The resulting radioactivity in the ipsilateral olfactory bulb was significantly higher than that in the contralateral bulb and peaked at 4 h. Tape section autoradiography showed that the radioactivity was concentrated in the olfactory nerve layer and the glomerular layer of the olfactory bulb. The olfactory transfer of dopamine was also studied <i>in vitro</i>. At a lower donor concentration, the mucosal-to-serosal dopamine permeability was higher than the serosal-to-mucosal permeability, but at a higher concentration, the permeability coefficients were similar. Together, these results suggest that the olfactory transfer of dopamine has an active component.</p><p>Olfactory transfer of fluorescein-labeled dextran through the epithelium and deeper tissues was studied in a rat model, which enabled visualization of the transfer using fluorescence microscopy. Although the epithelial transfer appeared to be mainly intracellular, transfer in the following deeper tissues was extracellular. Without altering the route of uptake, a gellan gum formulation enhanced the uptake of fluorescein dextran. The enhancing effect was considered likely to be the result of an increased residence time in the nasal cavity.</p><p>In conclusion, dopamine and fluorescein-labeled dextran were identified as suitable model compounds for the study of olfactory drug transfer mechanisms and the influence of drug formulation. Two new <i>in vitro</i> models of olfactory transfer were compared. Also, a rat model, which enabled the visualization of the entire nose-to-brain transfer, was developed.</p>

The blood-brain barrier restricts the access of many compounds, including therapeutic agents, to the brain. Several human studies indicate that nasal administration of hydrophilic compounds, such as peptides, can bypass the blood-brain barrier. The aims of this thesis were to develop and refine models for this direct nose-to-brain transfer. In a mouse model, [3H]-dopamine was given as a unilateral nasal dose. The resulting radioactivity in the ipsilateral olfactory bulb was significantly higher than that in the contralateral bulb and peaked at 4 h. Tape section autoradiography showed that the radioactivity was concentrated in the olfactory nerve layer and the glomerular layer of the olfactory bulb. The olfactory transfer of dopamine was also studied in vitro. At a lower donor concentration, the mucosal-to-serosal dopamine permeability was higher than the serosal-to-mucosal permeability, but at a higher concentration, the permeability coefficients were similar. Together, these results suggest that the olfactory transfer of dopamine has an active component. Olfactory transfer of fluorescein-labeled dextran through the epithelium and deeper tissues was studied in a rat model, which enabled visualization of the transfer using fluorescence microscopy. Although the epithelial transfer appeared to be mainly intracellular, transfer in the following deeper tissues was extracellular. Without altering the route of uptake, a gellan gum formulation enhanced the uptake of fluorescein dextran. The enhancing effect was considered likely to be the result of an increased residence time in the nasal cavity. In conclusion, dopamine and fluorescein-labeled dextran were identified as suitable model compounds for the study of olfactory drug transfer mechanisms and the influence of drug formulation. Two new in vitro models of olfactory transfer were compared. Also, a rat model, which enabled the visualization of the entire nose-to-brain transfer, was developed.

Given that Streptococcus pneumoniae can cause life-threatening pulmonary and systemic infection, an apparent paradox is that the bacterium resides, usually harmlessly, in the nasopharynx of many people. Humoral immunity is thought to be the primary defense against serious pneumococcal infection, and we hypothesized that nasopharyngeal colonization of mice results in the generation of an antibody response that provides long-term protection against lung infection. We found that survival of of C57L/6 mice after intranasal inoculation with wild-type serotype 4 strain TIGR4 pneumococci required B cells but not T cells, suggesting that nasopharyngeal colonization elicited a protective humoral immune response. In fact, intranasal inoculation resulted in detectable pneumococcal-specific antibody responses, and protected mice against a subsequent high-dose S. pneumoniae pulmonary challenge. B cells were required for this response, and transfer of immune sera from i.n. colonized mice, or monoclonal antibodies against phosphorylcholine, a common surface antigen of S. pneumoniae, was sufficient to confer protection. IgA, which is thought to participate in mucosal immunity, contributed to but was not absolutely required for protection from pulmonary challenge. Protection induced by i.n. colonization lasted at least ten weeks. Although it was partially dependent on T cells, depletion of CD4+ T cells at the time of challenge did not alter protection, suggesting that T cells did not provide essential help in activation of conventional memory cells. Peritoneal B1b cells and radiation-resistant, long-lived antibody secreting cells have previously been shown to secrete anti-pneumococcal antibodies and mediate protection against systemic infection following immunization with killed bacteria or capsular polysaccharide [1, 2]. We found that peritoneal cells were not sufficient for colonization-induced protection, but sub-lethally irradiated mice largely survived pulmonary challenge. Thus, our results are consistent with the hypothesis that nasopharyngeal colonization, a common occurrence in humans, is capable of eliciting extended protection against invasive pneumococcal disease by generating long-lived antibody-secreting cells.

Untersuchung zur Wirkung von intranasal verabreichtem Xylometazolin bei brachy- und normozephalen Hunden: Impuls-Oszillometrie und akustische Rhinometrie P. Franco, J. P. Hueber, G. U. Oechtering Klinik für Kleintiere, Arbeitsgruppe Brachyzephalie, Universität Leipzig Einleitung: Die zuchtbedingte Verkürzung des Gesichtsschädels brachy - zephaler Hunderassen hat zu einer Reihe klinisch relevanter morphologischer und physiologischer Veränderungen geführt. Hierzu zählen stenotische Naseneingänge, eine durch fehlgebildete Conchen obstruierte Nasenhöhle sowie ein verdicktes und verlängertes Gaumensegel. Diese Einengung der oberen Atemwege führt zu Atemnot und ausgeprägter Belastungsintoleranz, was als Brachyzephales Atemnotsyndrom (BAS) bezeichnet wird. Xylometazolin ist ein für den Menschen zugelassenes Alpha-Sympathomime - tikum. Topische und systemische Sympathomimetika werden beim Menschen zur Behandlung nasaler Kongestionen zum Abschwellen der Nasenschleimhaut eingesetzt. Zielstellung: Ausgehend von der Hypothese, dass Xylometazolin auch bei Hunden zu einem Abschwellen der Nasenschleimhaut führt und dass die Morphologie der Nasenmuscheln sich zwischen den zwei Zielgruppen unterscheidet, sollte der intranasale Strömungswiderstand bei brachy- und normozephalen Hunden vor und nach Xylometazolingabe mit Impuls- Oszillometrie untersucht werden. Zusätzlich wurden in der Beaglegruppe mit akustischer Rhinometrie das Nasenhöhlenvolumen und die minimale Querschnittsfläche bestimmt. Methodik: Die Messungen erfolgten nach dem Prinzip der Impuls-Oszillometrie (IOS) und akustischer Rhinometrie. In einer prospektiven klini - schen Studie wurden 10 brachyzephale Hunde (5 Möpse, 5 Französische Bulldoggen) und 6 Beagles untersucht. Bei den anästhesierten, spontan atmenden Tieren wurde der intranasale Strömungswiderstand mit Impuls- Oszillometrie unmittelbar vor und 30 Minuten nach intranasal verabreichtem Xylometazolin gemessen. Messungen mit akustischer Rhinometrie erfolgten zusätzlich vor und nach Xylometazolingabe. Bei allen Tieren wurden die oberen Atemwege endoskopisch und computertomographisch untersucht. Ergebnisse: Der aus drei Messungen gemittelte intranasale Strömungswiderstand bei Patienten mit BAS betrug vor Xylometazolingabe 0,87 ± 0,097 kPa/(L/s). Nach Xylometazolingabe reduzierte sich der intranasale Strömungswiderstand um etwa 48% auf 0,42 ± 0,55 kPa/(L/s). Vergleich - bare Ergebnisse ermittelten wir in der Beaglegruppe. Nach Xylometazolingabe ergab sich bei den Beagles eine Zunahme des Nasenvolumens und der minimalen Querschnittsfläche. Schlussfolgerungen: Wir konnten erstmals zeigen, dass Xylometazolin auch bei Hunden zum Abschwellen der Nasenschleimhaut führt und sich das Abschwellverhalten zwischen brachy- und normozephalen Hunden kaum voneinander unterscheidet. A13 Abstracts 18. Jahrestagung der DVG-FG InnLab © Schattauer 2010 Tierärztliche Praxis Kleintiere 1/2010