The effect of absorbable packing on the healing of nasal respiratory epthelium after endoscopic sinus surgery (ESS) was examined in a diseased sheep model. Full thickness injuries were created on the lateral nasal wall of sheep infested with Oestrus ovi. Sites of injury were packed on one side with hyaluronic acid (HA) packing or hyaluronic acid packing impregnated with insulin-like growth factor- 1 (HA+IGF1) in a randomized fashion. The opposite side was left unpacked as a control. Biopsies were obtained for light microscopy, scanning electron microscopy, and ciliary beat frequency (CBF) analysis over a period of 16 weeks. Statistical analysis of results was performed in order to determine if any intervention had any impact on healing and to determine if there was any correlation between extent of regeneration as assessed by electron microscopy and CBF. Furthermore assessment of the effect of isotonic and hypertonic saline on ciliary beat frequency was performed in healthy human volunteers. Reepithelialization was increased in the HA+IGF1 group compared to the HA group and controls at eight weeks after injury but not at later time points. Cilial regeneration was improved in the HA+IGF1 group compared to the HA group and controls at 16 weeks. CBF was noted to be worse at the eight week time point with the HA+IGF1 group compared to the HA group and controls, but no other statistically significant effects on CBF were noted. This most likely represents a spurious finding. Wide distributions of CBF results were noted, reflecting numerous missing data points due to methodological difficulties. There was a trend noted toward increased CBF with improved grades of reciliation, although this correlation was not statistically significant. However this trend was supported by the finding of statistically significant differences between individual and combined grades of reciliation. Hypertonic saline was found to have a ciliostimulatory effect when compared to normal saline at 5 minutes after administration in healthy human subjects. This effect had disappeared by 60 minutes after administration. It is suggested that the presence of insulin-like growth factor- 1 at the time of mucosal injury improves epithelial regeneration in the short term, but is not sufficient for this effect to be sustained. This improved early epithelial regeneration forms a foundation for cilial regeneration, as is reflected in an improved grade of reciliation at 16 weeks. Our interventions had no effect on CBF, and various experimental problems made it difficult to provide further comment on CBF results. There is evidence that CBF improves as the grade of cilial regeneration improves following ESS. Furthermore, hypertonic saline appears to also have a positive impact on CBF, which is likely to reflect changes in the rheological properties of mucous. A number of possible avenues of enquiry are delineated and recommendations for future research are outlined. / Thesis (M.S.)--Department of Surgery, 2005.

Les vaccins des muqueuses s'administrent facilement et favorisent une immunité humorale et cellulaire au niveau des muqueuses. Ainsi, ils offrent une protection optimale contre les pathogènes envahissant par les muqueuses. Cependant, la mise sur le marché de nouveaux vaccins des muqueuses est entravée par le manque d'adjuvants des muqueuses efficaces et sans effets secondaires. La toxine cholérique (CT) est une entérotoxine à fort pouvoir adjuvant mais sa toxicité empêche son utilisation chez l'homme. En tant que modèle expérimental, elle a permis de comprendre l'importance de l'activité enzymatique et des récepteurs dans l'adjuvanticité. Notre travail se base sur l'observation que des doses sublétales de la toxine œdémateuse de Bacillus anthracis (EdTx) n'inhibent pas la réponse immune à des vaccins « nasaux » contenant CT comme adjuvant. Nous avons montré que les dérivés EdTx représentent une nouvelle classe d'adjuvants qui donnent des réponses systémiques et des muqueuses à des protéines vaccinales administrées par de multiples voies, notamment nasale. Contrairement à CT qui se fixe aux gangliosides, EdTx se fixe aux récepteurs des toxines de l'anthrax et ne cible pas les tissus du système nerveux central après administration nasale. Le facteur inné nerve growth factor intervient dans les réponses des muqueuses induites par CT mais n'affecte pas l'adjuvanticité d'EdTx in vivo. L'activité adjuvante d'EdTx implique aussi l'augmentation des fonctions de présentation de l'antigène. Enfin, nous avons montré qu'EdTx est un adjuvant efficace par les voies transcutanée et sublinguale, bien que les IgA des muqueuses ne soient induits qu'après immunisation sublinguale.

<p>The nasal administration of drugs offers advantages over administration by intravenous injection. Drugs can be rapidly absorbed through the nasal mucosa, resulting in a rapid onset of action, and also avoiding degradation in the gastrointestinal tract and first-pass metabolism in the liver. The olfactory receptor cells, which are in direct contact with both the environment and the central nervous system (CNS), are potential routes for drugs into the CNS. The olfactory pathway thus circumvents the blood brain barrier (BBB) which prevents many systemically administered drugs from entering the brain.</p><p>The studies used compounds active in the CNS and the experiments were performed in rodents. The nasal bioavailability of (S)-UH-301, NXX-066 and [³H]-dopamine was investigated in a rat model; uptake into the cerebrospinal fluid (CSF) was compared after nasal and intravenous administration. The concentrations of S-UH-301 and NXX-066 in plasma and CSF were measured with high performance liquid chromatography. The possible transfer of dopamine and neurotensin along the olfactory pathway after nasal administration to mice was studied using brain tissue sampling and autoradiography. The radioactivity content in blood, CSF and dissected brain tissue samples after administration of [³H]-dopamine and [³H]-neurotensin was assessed using liquid scintillation, and thin layer chromatography (TLC) was used to investigate the metabolic fate of [³H]-dopamine.</p><p>The results of this thesis suggest that nasal administration of CNS-active compounds with low oral bioavailability is an interesting and workable alternative to intravenous injection. The small lipophilic compounds (S)-UH-301 and NXX-066 were rapidly and completely absorbed after nasal administration, although hard evidence of direct transfer from the nose remains elusive. Radioactivity measurements in the olfactory bulb following nasal administration of</p><p>[³H]-dopamine and [³H]-neurotensin indicate that transfer occurred. The TLC results showed the presence of unchanged dopamine in the olfactory bulb but it is less clear from initial results with neurotensin, which radioactive products of this molecule reached the olfactory bulb, and further studies are required.</p>

Background A perforation in the nasal septum can cause symptoms such as bleeding, obstruction, crusts and pain, and can be a challenge to treat. Surgery is the treatment of choice, but disease, size of the perforation, or the patient’s wish may contradict surgery. A custom-made silicone obturator is a successful treatment option, but little is known how this treatment affects the microbial flora. The purposes of this thesis were (i) to investigate the microbial flora around symptomatic nasal septal perforations before treatment, (ii) during and after a 12-month treatment period with a custom-made obturator, (iii) to compare the microbial flora around symptomatic perforations with the flora from the same area of the septum in healthy individuals, (iv) to investigate the microbial colonization of the silicone obturator, and (v) also to investigate the water sorption, solubility and if the wettability of silicones are affected by water. The hypotheses were (i) that the bacterial flora around symptomatic perforations would not differ from that found in healthy individuals, apart from a possible presence of Helicobacter pylori; (ii) the bacterial flora would change in composition during the course of treatment and that microorganisms and proteins could be seen on the surface of the silicone obturators; (iii) a material that has adsorbed water would also show an increase in wettability and the surface free energy of the material.  Methods Twenty-seven patients and 101 healthy individuals volunteered. Swabs were made around the rim of the perforation, or on the septum in the locus Kisselbachi area in the healthy individuals. Bacteria and fungi were isolated and identified with standard laboratory techniques. A biopsy of the granulated tissue at the perforation was taken and cultivated for Helicobacter pylori. Swabs were also taken three, six and twelve months after inserting the obturator. The obturator was analysed after being used twelve months in the nose.  Seven silicones were tested for water sorption and solubility according to ISO standards 1567:1999 and ISO 10477:2004. The change in wettability was examined by measuring the contact angle with a contact goniometer at various stages of the sorption/solubility test. Results Staphylococcus aureus was present in 88% of the untreated patients. With treatment a significant reduction of S. aureus occurred to 54.5% (p&lt;0.05). In the healthy group S. aureus was present in 13% of the subjects. No Helicobacter pylori could be cultivated from the biopsies taken of the granulated tissue at the perforation. The flora round the untreated perforation was dominated by S. aureus with few other bacterial species detected. In the healthy group there was a diversified flora with both aerobic and anaerobic bacteria. SEM revealed a rough surface on the silicone obturator and crazing of the silicone surrounding the pigment granules. Both bacteria and proteins could be seen on the obturators in SEM. Candida albicans was detected in one obturator, but not in the mucosal swab at the corresponding time. That patient had, however, been treated for Candida in the nose six months prior to the last visit in the study. Wettability was affected but did not increase with amount of adsorbed water. Some materials showed an increase and some a decrease in the surface-free energy. The tested addition silicones showed little sorption and solubility. Conclusions The patients with symptomatic perforations of the nasal septum had a bacterial flora totally dominated by S. aureus. The massive presence of S. aureus around symptomatic perforations may have an impact on the persistence of the granulated and inflamed tissue present in symptomatic perforations, thus forming a vicious circle with bleeding and crustation. S. aureus dominance in the mucosa surrounding symptomatic perforations was diminished by using a custom-made obturator. The microbial flora became more diversified with the treatment, although not resembling the flora in healthy individuals. The microbial flora of the obturators was similar, but not the same as the corresponding mucosal flora. The discovery of Candida in the obturator of a patient who had been treated for Candida in the nose six months earlier suggests that obturators need to be exchanged when fungal infections are being treated to prevent the fungus from re-infecting the patient at a later stage. The silicone had a rough surface and a poor wettability, both aspects favours colonization of microorganisms. The silicone was negatively affected by the colouring pigments, this should be considered when colouring is not necessary. The slight, but existing solubility of silicones emphasises the importance of using medical grade silicones that are more purified than industrial silicones.

The nasal administration of drugs offers advantages over administration by intravenous injection. Drugs can be rapidly absorbed through the nasal mucosa, resulting in a rapid onset of action, and also avoiding degradation in the gastrointestinal tract and first-pass metabolism in the liver. The olfactory receptor cells, which are in direct contact with both the environment and the central nervous system (CNS), are potential routes for drugs into the CNS. The olfactory pathway thus circumvents the blood brain barrier (BBB) which prevents many systemically administered drugs from entering the brain. The studies used compounds active in the CNS and the experiments were performed in rodents. The nasal bioavailability of (S)-UH-301, NXX-066 and [3H]-dopamine was investigated in a rat model; uptake into the cerebrospinal fluid (CSF) was compared after nasal and intravenous administration. The concentrations of S-UH-301 and NXX-066 in plasma and CSF were measured with high performance liquid chromatography. The possible transfer of dopamine and neurotensin along the olfactory pathway after nasal administration to mice was studied using brain tissue sampling and autoradiography. The radioactivity content in blood, CSF and dissected brain tissue samples after administration of [3H]-dopamine and [3H]-neurotensin was assessed using liquid scintillation, and thin layer chromatography (TLC) was used to investigate the metabolic fate of [3H]-dopamine. The results of this thesis suggest that nasal administration of CNS-active compounds with low oral bioavailability is an interesting and workable alternative to intravenous injection. The small lipophilic compounds (S)-UH-301 and NXX-066 were rapidly and completely absorbed after nasal administration, although hard evidence of direct transfer from the nose remains elusive. Radioactivity measurements in the olfactory bulb following nasal administration of [3H]-dopamine and [3H]-neurotensin indicate that transfer occurred. The TLC results showed the presence of unchanged dopamine in the olfactory bulb but it is less clear from initial results with neurotensin, which radioactive products of this molecule reached the olfactory bulb, and further studies are required.

<p>Although monophosphoryl lipid A was recently approved by the Food and Drug Administration, more vaccine adjuvants are needed to meet the demand for vaccines against new, emerging, and re-emerging diseases. Additionally, characterizing the mechanisms of action of potent vaccine adjuvants is important for moving toward more rational vaccine design based on the careful selection of antigens and adjuvants to stimulate only the desired immune responses. Two experimental vaccine adjuvants, compound 48/80 (C48/80) and IL-1, were evaluated in these studies. The safety and efficacy of the mast cell activator C48/80 was evaluated when used as an adjuvant delivered intradermally (ID) with recombinant anthrax protective antigen (rPA) in comparison with two well-known adjuvants. Mice were vaccinated in the ear pinnae with rPA or rPA + C48/80, CpG oligodeoxynucleotides (CpG), or cholera toxin (CT). All adjuvants induced similar increases in serum anti-rPA IgG and lethal toxin-neutralizing antibodies. C48/80 induced balanced cytokine production (Th1/Th2/Th17) by antigen-restimulated splenocytes, minimal injection site inflammation, and no antigen-specific IgE. Our data demonstrate that C48/80 is a safe and effective adjuvant, when used by the intradermal route, to induce protective antibody and balanced Th1/Th2/Th17 responses. Histological analysis demonstrated that vaccination with C48/80 reduced the number of resident mast cells and induced an injection-site neutrophil influx within 24 hours. Nonetheless, rPA + C48/80 significantly increased antigen-specific IgG titers in mast cell-deficient mice compared to antigen alone, suggesting that C48/80 has mast cell-dependent and mast cell-independent mechanisms of action.</p><p>IL-1alpha and beta have been shown to have strong mucosal adjuvant activities, but little is known about their mechanism of action. Bone marrow chimeric mice were intranasally vaccinated with Bacillus anthracis lethal factor (LF) with or without 4 µg IL-1alpha or a control adjuvant (cholera toxin) to determine if IL-1R1 expression on stromal cells or hematopoietic cells was sufficient for the maximal adjuvant activity of nasally delivered IL-1alpha. IL-1alpha was not active in IL-1R1-deficient (<italic>Il1r1</italic>-/-) mice given <italic>Il1r1</italic>-/- bone marrow, demonstrating that the adjuvant activity of IL-1 was due to the presence of IL-1R1 and not contaminants. Cytokine and chemokine responses induced by vaccination with IL-1alpha were predominantly derived from the stromal cell compartment and included G-CSF, IL-6, IL-13, MCP-1, and KC. Nasal vaccination of <italic>Il1r1</italic>-/- mice given wild-type bone marrow (WT-->KO) and WT-->WT mice with LF + IL-1alpha induced maximal adaptive immune responses, while vaccination of wild-type mice given <italic>Il1r1</italic>-/- bone marrow (KO-->WT) mice resulted in significantly decreased production of LF-specific serum IgG, IgG subclasses, lethal toxin-neutralizing antibodies, and mucosal IgA compared to WT-->KO and WT-->WT mice (p < 0.05). Our results suggest that IL-1R1 expression in the hematopoietic compartment is sufficient for the maximal induction of antigen-specific adaptive immunity after nasal vaccination adjuvanted with IL-1alpha and that while stromal cells are required for maximal adjuvant-induced cytokine production, the adjuvant-induced stromal cell cytokine responses are not required for effective induction of adaptive immunity.</p> / Dissertation

Ce travail a pour objectif la caractérisation articulatori-acoustique de la nasalité: nature des mouvements du port vélopharyngé, caractéristiques acoustiques liées. La construction d'un modèle articulatoire linéaire 3D monosujet du conduit nasal à partir d'images IRM et CT a fait émerger deux degrés de liberté parmi les mouvements du voile du palais et de la paroi nasopharyngée. Le premier, prédominant, correspond à un mouvement conjoint vertical oblique du voile et horizontal de la paroi pharyngée, traduisant l'effet de sphincter du port vélopharyngé, et le second à un petit mouvement horizontal du voile seul, modifiant sensiblement l'aire de couplage nasal. L'espace des mouvements du modèle décrit exactement celui d'un point du voile obtenu par articulographie électromagnétique. Les fonctions d'aire réalistes du conduit nasal déduites du modèle ont permis de déterminer les fonctions de transfert acoustiques de sept voyelles et l'influence acoustique des mouvements du voile.

Cleft palate is one of the most common congenital malformations in humans which occurs at a frequency of approximately 1:700 live births worldwide. Sine Oculis-related homeobox 2 (Six2) is a member of the vertebrate Six gene family that encode proteins that are transcription factors. Six2 has been reported to be a downstream target of Homeobox a2 (Hoxa2), a gene that plays a direct a role in mouse secondary palate (SP) development. In my thesis, I utilized quantitative real time Polymerase Chain Reaction (qPCR), Western blot analysis and fluorescence immunohistochemisrty (IHC) to characterize the spatial and temporal distribution patterns of Six2 in the developing SP. Additionally, I also employed in vivo cell counting analysis and in vitro cell proliferation assays to investigate the role of Six2 during palate mesenchymal cell proliferation. My study examined the temporal and spatial distribution of Six2 in the developing mouse palatal mesenchyme and epithelia in both wild-type and Hoxa2 null mice. Six2 was expressed throughout the period of embryonic palatogenesis, with the highest levels of Six2 mRNA and protein observed in palatal shelves at E13.5 in both wild-type and Hoxa2 null mice. Six2 protein expression at all stages of SP development (E12.5 to E15.5) increased in the anterior to posterior (A-P) direction with highest expression in the posterior regions of the developing SP. In addition, expression of Six2 protein was higher in the oral half of the palatal mesenchyme compared to the nasal half of the palatal mesenchyme. Interestingly, Six2 protein was expressed in the nasal palatal epithelium but was completely absent from the oral palatal epithelium. Loss of the Hoxa2 gene induced up regulation of Six2 protein and mRNA in the developing palate across all stages of palatogenesis. In the Hoxa2 null mice, there was a significant increase in cell proliferation (Ki-67 positive cells) and the percentage of actively proliferating cells that were co-expressing Six2 protein (Six2/Ki-67 double positive cells) along both the A-P and oral-nasal (O-N) axes of the developing SP. Also, the highest percentage of actively proliferating cells and Six2/Ki-67 double positive cells was observed in the nasal half of the posterior palatal mesenchyme. Furthermore, Six2 siRNA knock down in mouse embryonic palatal mesenchyme (MEPM) cell cultures restored cell proliferation and Cyclin D1 expression in the Hoxa2 null cell cultures to wild-type levels. Collectively, my data reveals a novel spatial and temporal expression profile for Six2 in the developing mouse SP and the potential role it might play during the epithelial-mesenchymal cross talk that drives palatal shelf cell proliferation and out growth.

Approximately 350 million people worldwide suffer from diabetes mellitus (DM) and this number increases yearly. Since the discovery and clinical application of insulin in 1921, subcutaneous injections have been the standard treatment for DM. Because insulin is hydrophilic and has a high molecular weight and low bioavailability, this molecule is poorly absorbed if administered orally. The aim of this study is to evaluate nasal delivery systems for insulin, using Sprague Dawley rats as the nasal absorption model. Pheroid technology and N-trimethyl chitosan chloride (TMC) with different dosages of insulin (4, 8 and 12 IU/kg bodyweight insulin) was administered in the left nostril of the rat by using a micropipette. Pheroid technology is a patented (North-West University) carrier system consisting of a unique oil/water emulsion that actively transports drug actives through various physiological barriers. These formulations were administered nasally to rats in a volume of 100 p/kg bodyweight in different types of Pheroids (vesicles, with a size of 1.7 1 - 1.94 pm and microsponges, with a size of 5.7 1 - 8.25 pm). The systemic absorption of insulin was monitored by measuring arterial blood glucose levels over a period of 3 hours. The TMC formulation with 4 IU/kg insulin produced clinically relevant levels of insulin in the blood and as a result also the maximal hypoglycaemic effect. TMC is a quaternary derivative of chitosan and is able to enhance the absorption of various peptide drugs by opening tight junctions between epithelial cells. Pheroid formulations were also effective in lowering blood glucose levels but only at higher doses (8 and 12 IU/kg) of insulin. This study indicated that Pheroid rnicrosponges had a faster onset of action and a slightly better absorption of insulin when compared to Pheroid vesicles, but many more studies are needed in this field. Although the results of this study with absorption enhancers are encouraging, nasal insulin bioavailability is still very low, and the Pheroid formulations and long-term safety of nasal insulin therapy have yet to be investigated. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2007.

Advances in biotechnology and recombinant technologies have lead to the production of several classes of new drugs such as peptide and protein drugs. These compounds are mostly indicated for chronic use but their inherent characteristics such as size, polarity and stability prevent them from incorporation in novel dosage forms. The bioavailability of nearly all peptide drugs is very low due to poor absorption from the administration site. Several challenges confront the pharmaceutical scientist in developing effective and innovative dosage forms for these classes of drugs. A lot of attention has been given to the nasal route of drug administration for delivery of peptide drugs. The availability of several promising classes of absorption enhancers and new drug delivery technologies has also prompt scientists to develop new delivery systems for nasal administration of peptide drugs. It has been shown in recent years that N-trimethyl chitosan chloride (TMC), a quaternary derivative of chitosan, is effective in enhancing the absorption of several peptide drugs, both in the peroral route and in the nasal route of drug administration. Early indications are that new drug delivery technologies such as Pheroid technology will also be able to enhance peptide drug absorption in the nasal route. The aim of this study was to evaluate and compare the absorption enhancing abilities of TMC and Pheroid technology in the nasal delivery of calcitonin, a peptide hormone with low bioavailability. Pheroid vesicles and Pheroid microsponges were prepared and characterized for their morphology and size distribution. Calcitonin was entrapped into these vesicles and microsponges and TMC and TMO solutions (0.5 % w/v), containing calcitonin, was also prepared. These formulations were administered nasally to rats in a volume of 100 μl/kg body-weight to obtain a final concentration of 10 IU/kg body-weight of calcitonin. Plasma calcitonin and calcium levels were determined over a period of 3 hours. The results of this study clearly indicated that both Pheroid formulations and the TMC formulation increase the nasal absorption of calcitonin with a resulting decrease in plasma calcium levels, indicating an increased absorption of calcitonin. The highest increase in calcitonin absorption was obtained with the TMC formulation and this was explained by the difference in the mechanism of action in enhancing peptide absorption between TMC and Pheroid technology. It was concluded that Pheroid technology is also a potent system to enhance peptide drug delivery and that the exact mechanism of action should be investigated further. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2006.