Staphylococcus is a significant cause of human infection and mortality, worldwide. Currently, there are greater than 60 taxa within Staphylococcus, and nearly all are pathogenic. The collective potential for virulence among species of Staphylococcus heightens the overall clinical significance of this genus and argues for a thorough understanding of the evolutionary relationships among species. Within Staphylococcus, aureus is the most common cause of human infection, where nasal carriage of this bacterium is a known risk factor for autoinfection. The predisposition to infection by nasal carriers of S. aureus, and the ease with which strains are transferred between individuals, suggests that nasal carriage is a major vector for the transmission of virulent strains throughout the community. This hypothesis, however, has not been assessed in any great detail to identify the genetic relationships between clinical isolates of S. aureus and those strains being carried asymptomatically throughout the community. Also lacking within this field is a unified and robust estimate of phylogeny among species of Staphylococcus. Here, we report on a highly unified species phylogeny for Staphylococcus that has been derived using multilocus nucleotide data under multiple Bayesian and maximum likelihood approaches. Our findings are in general agreement with previous reports of the staphylococcal phylogeny, although we identify multiple previously unreported relationships. Regardless of methodology, strong nodal support and high topological agreement was observed with only minor variations in results between methods. Based on our phylogenetic estimates, we propose that Staphylococcus species can be evolutionarily clustered into 15 groups, and six species groups. In addition, our more defined phylogenetic analyses of S. aureus revealed strong genetic associations between both nasal carriage strains and clinical isolates. Genetic analyses of hypervariable regions from virulence genes revealed that not only do clinically relevant strains belong to identical genetic lineages as the nasal carriage isolates, but they also exhibited 100% sequence similarity within these regions. Our findings indicate that strains of S. aureus being carried asymptomatically throughout the community via nasal colonization are genetically related to those responsible for high levels of infection and mortality. Due to nasal carriage of S. aureus being a risk factor for autoinfection, standardized preoperative decolonization has become a major consideration for the prevention of nosocomial infection. Toward this end, we have identified the macrocyclic ?-defensin analogue RC-101 as a promising anti-S. aureus agent for nasal decolonization. RC-101 exhibited bactericidal effects against S. aureus in both epithelium-free systems, and ex vivo models containing human airway epithelia. Importantly, RC-101 exhibited potent anti-S. aureus activities against all strains tested, including USA300. Moreover, RC-101 significantly reduced the adherence, survival, and proliferation of S. aureus on human airway epithelia without any noted cellular toxicity or the induction of a proinflammatory response. Collectively, our findings identify RC-101 as a potential preventative of S. aureus nasal colonization.

Thesis (Ph. D.)--West Virginia University, 2003. / Title from document title page. Document formatted into pages; contains xiii, 82, [148] p. : ill. (some col.). Includes abstract. Includes bibliographical references.

Thesis (Ph.D. in Neuroscience) -- University of Colorado Denver, 2007. / Typescript. Includes bibliographical references (leaves 129-151). Free to UCD affiliates. Online version available via ProQuest Digital Dissertations;

Introdução: A patogenia da rinossinusite crônica com polipose nasossinusal não está completamente estabelecida e existem algumas explicações para essa doença como os superantigenos, o desequilíbrio inflamatório e, mais recentemente, o biofilme. Objetivos: Avaliar a associação entre a presença do biofilme e a presença de rinossinusite crônica com polipose nasossinusal. Avaliar o quadro clínico e radiológico pré-operatória e pós-operatória segundo a presença do biofilme. Métodos: Este é uma estudo realizado em um hospital terciário universitário. A primeira parte foi um estudo caso-controle com um grupo de 33 pacientes consecutivos com rinossinusite crônica com polipose nasossinusal submetidos a cirurgica endoscópica nasossinusal e um grupo controle de 27 pacientes submetidos a septoplastia para tratamento de obstrução nasal. As amostras da mucosa foram coletadas no intra-operatório para avaliação por microscopia eletrônica de varredura para determinar a presença do biofilme. A segunda parte foi um estudo prospectivo em que dados pré-operatórios e pós-operatórios foram registrados, incluindo avaliações padronizadas da qualidade de vida doença-específica relacionadas à obstrução nasal e à rinossinusite, da endoscopia nasal e da tomografia de cavidades paranasais. A análise estatísca foi realizada. Para todos os testes um p=0.05 foi considerado significativo. Resultados: Os biofilmes foram encontrados em 72.7% (24/33) dos pacientes com rinossinusite crônica com polipose nasossinusal e 48.1% (13/27) dos pacientes submetidos a septoplastia (Odd ratio=2.87, IC95% 0.9796-8.419, p=0.051). Este foi o primeiro estudo a analisar o efeito da presença do biofilme nos resultados pós-operatórios com medidas padronizadas de um grupo de pacientes apenas com rinossinusite crônica com polipose nasossinusal. O biofilme estava presente em 72.4% (21/29) dos pacientes que completaram o seguimento. Os pacientes com biofilmes apresentaram uma pior pontuação pré-operatória NOSE e Lund-Kennedy estatísticamente significativos, mas uma mediana semelhante na pontuação total do SNOT-20. Os pacientes com biofilme apresentaram uma melhor resultado na pontuação Lund-Kennedy (p=0.036). Estes pacientes apresentaram piores resultados no SNOT-20 e resultados similares quanto ao NOSE e o Lund-Mackay. Conclusão: Os biofilmes foram demonstrados presentes nos pacientes submetidos a cirurgia endoscópica funcional para rinossinusite crônica com polipose nasossinusal mas também nos controles. Embora a prevalência não tenha sido diferente significativamente, o intervalo de confiança extremamente amplo de 95%, que apenas cruza a unidade, sugere que uma diferença significativa pode ter sido perdida por causa do baixo poder estatístico e estudos futuros serão necessários. Os biofilmes estiveram relacionados com pior qualidade de vida doença-específica pré-operatória NOSE e avaliação endoscópica (Lund-Kennedy), e melhores resultados endoscópicos. Nossos resultados sugerem que nos pacientes com uma melhora clínica significativa após a cirurgia, o biofilme representou um papel mais predominante na fisiopatologia da doença. Neste subgrupo, a cirurgia provavelmente removeu a quantidade de biofilme necessária para restaurar o desequilíbrio inflamatório na mucosa / Introduction: The pathogenesis of chronic rhinosinusitis with nasal polyps is not completely established and there are some explanations for this disease, such as superantigens, inflammatory imbalance and, more recently, biofilms. Objective: Evaluate the association of biofilms presence and chronic rhinosinusitis with nasal polyps. Evaluate outcomes after sinus surgery for chronic rhinosinusitis with nasal polyps according to the presence of biofilms. Methods: This is a University based-tertiary care center study. The first part was a case-control study that evaluated a group of 33 consecutive patients undergoing functional endoscopic sinus surgery for chronic rhinosinusitis with nasal polyps and a control group of 27 patients undergoing septoplasty for nasal obstruction treatment. Mucosal samples were harvested intra-operatively for scanning electron microscopic examination to determine biofilms presence. The second part was a prospective study. Preoperative and follow up data were recorded, including standardized evaluations of disease-specific quality of life related to nasal obstruction and rhinosinusitis, of nasal endoscopy and sinus computer tomography scan. Statistical analysis was performed. For all statistical tests p=0.05 was considered significant. Results: Biofilms were found in 72.7% (24/33) of chronic rhinosinusitis with nasal polyps patients and in 48.1%(13/27) of septoplasty patients (Odds ratio = 2.87, CI95% from 0.9796 to 8.419, p=0.051). This was the first report to analyze the effect of biofilms in outcomes with standardized measures of a group of only chronic rhinosinusitis with nasal polyps patients. Biofilms were present in 72.4% (21/29) of these patients. Patients with biofilms had a statistically significant worst preoperative score related to nasal obstruction and nasal endoscopy, but a similar median sinusitis total score. Patients with biofilms presented better Lund-Kennedy outcome (-3[5]vs.-1[2],U=46.0,p=0.036), but the best endoscopic improvement might reflect the worst clinical preoperative status. These patients had worst outcomes in SNOT-20 (-0.75[1.15]vs.-1.30[1.32],U=69.0,p=0.21) and similar outcomes in NOSE(-55.0[50.0] vs. -60.0[50.0], U=81.0,p=0.67) and Lund-Mackay (-4[5]vs.-4[4]),U=75.5,p=0.49). Patients with biofilms presented better Lund-Kennedy outcome (p=0.036). There was a correlation among some QoL outcome scores in both groups. Conclusion: Biofilms were demonstrated to be present in patients undergoing functional endoscopic sinus surgery for chronic rhinosinusitis with nasal polyps but also in controls. Although the prevalence was not significantly different, the extremely wide 95% confidence interval, which just crosses unity, suggests that a meaningful clinical difference may have been missed because of low statistical power and that further study is necessary. Biofilms were related with worst preoperative disease-specific quality of life questionnaire (NOSE) and endoscopic evaluation (Lund-Kennedy), and better endoscopic outcome. Our findings suggest that in patients with a significant clinical improvement after surgery, the biofilm had a more predominant role in the pathophysiology of the disease. In this subgroup, the surgery probably removed the amount of biofilms needed to restore the mucosal inflammatory imbalance

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.

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<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 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.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Linköping : Univ., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2008. / Härtill 5 uppsatser.