Background: Azithromycin polarizes macrophages into an alternative phenotype and promotes a regulated immunity. Arginase is an important effector of these macrophages believed to play an essential role in decreasing injury and promoting repair. Hypothesis: Decreases in inflammation in response to Pseudomonas aeruginosa (PA) pneumonia achieved by polarizing macrophages to an alternative phenotype is dependent upon the production of arginase. Methods: Requirement of arginase was examined by pharmacological inhibition using S-(2-boronoethyl)- l-cysteine (BEC) or l-norvaline and by infecting arginase-1 conditional knock-out mice (Arg1flox/flox;Lyz2-cre (Arg1Δm)) with PA intratracheally. Arg1ΔM and control Arg1flox/flox mice were then dosed with azithromycin daily via oral gavage beginning four days prior to infection. Analysis of weight loss in addition to characterization of inflammatory cells and cytokine production via flow cytometry was performed. Macrophages were then stimulated with LPS and polarized with IL4/13, IFNγ, or azithromycin plus IFNγ. Western blot for signaling mediators, p65 translocation assay, and immunofluorescence were performed. Results: Myeloid arginase-1 deletion resulted in greater morbidity along with more severe inflammatory response compared to the Arg1flox/flox mice. Arg1Δm mice had greater numbers of neutrophils, macrophages, and lymphocytes in their airways and lymph nodes compared to the Arg1flox/flox mice. Conversely, global arginase inhibition resulted in greater weight loss along with greater neutrophil and macrophage infiltration compared to Arg1Δm mice. BEC and l-norvaline treated mice had higher numbers of lymphocytes in their lymph nodes with variable effects on airway lymphocyte counts. Azithromycin treatment comparably reduced the acute inflammatory responses in both Arg1Δm and Arg1flox/flox mice. To evaluate this mechanism, we show in vitro that azithromycin decreases NF-κB activation by preventing p65 nuclear translocation and by decreasing STAT1 activation in a concentration-dependent manner. These effects were reversed with IKKβ inhibition. Conclusions: Myeloid arginase is essential for control of inflammatory responses in PA pneumonia with potentially different effects of other cellular sources demonstrated with global arginase inhibition. Azithromycin reduces excessive inflammation even in the absence of arginase, potentially through a cross-inhibitory mechanism involving STAT1 and NF-κB pathways through IKKβ.

Orientador: James Venturini / Resumo: Paracoccidioidomicose (PCM) é uma micose sistêmica causada por fungos do gênero Paracoccidioides; suas principais formas clínicas são aguda/subaguda, crônica e residual. A PCM é uma doença restrita a países da América Latina com maior incidência no Brasil, especialmente entre os trabalhadores rurais. A maioria dos pacientes com a forma crônica da doença, mesmo após tratamento eficaz, apresentam sequelas, incluindo fibrose pulmonar e adrenal. Os problemas sociais, econômicos e psicológicos desencadeados pela fibrose pulmonar são subestimados; além disso, a fibrose na PCM permanece negligenciada, uma vez que não há tratamento. Dessa forma, o estudo teve por objetivo investigar a influência de drogas com potencial antifibrótico (pentoxifilina - PTX, azitromicina - AZT e talidomida - Thal) associadas aos tratamentos antifúngicos com itraconazol - ITC e cotrimoxazol - CMX em modelo murino de PCM pulmonar. Para tanto, camundongos BALB/c machos foram inoculados com leveduras do isolado 326 de P. brasiliensis e após 8 semanas de infecção foi dado início aos esquemas terapêuticos: PTX/ITC, PTX/CMX, AZT/ITC, AZT/CMX, Thal/ITC e Thal/CMX. Após 8 semanas de tratamento, os animais foram eutanasiados a fim de se avaliar a deposição de fibras colágenas, produção de hidroxiprolina, recuperação de fungos viáveis e a porcentagem das áreas com lesão nos pulmões e peso corporal. Visando identificar os mecanismos envolvidos foi avaliada a produção de TGF-β1, CCL3, IFN-γ, TNF-α, IL-10, VEGF, IL-6,... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Paracoccidioidomycosis (PCM) is a systemic mycosis caused by fungi of Paracoccidioides genus; the main clinical forms are acute/subacute, chronic and residual. PCM is a disease restricted to Latin American countries with a higher incidence in Brazil, especially among rural workers. Most patients with the chronic form, even after effective treatment, present sequelae, including pulmonary and adrenal fibrosis. The social, economic and psychological problems triggered by pulmonary sequels are underestimated. In addition, fibrosis in PCM remains neglected, since there is no treatment. The aim of this study was to investigate the influence of antifibrotic drugs (pentoxifylline - PTX, azithromycin - AZT and thalidomide - Thal) associated with antifungal treatments with itraconazole - ITC and cotrimoxazole - CMX in a murine model of pulmonary PCM. Male BALB/c mice were inoculated with P. brasiliensis “isolated 326” and after 8 weeks of infection the treatment were started: PTX/ITC, PTX/CMX, AZT/ITC, AZT/CMX, Thal/ITC and Thal/CMX. After 8 weeks of treatment, the mice were euthanized in order to evaluate the deposition of collagen fibers, hydroxyproline production, recovery of viable fungi and the percentage of areas with injury in lung and body weight. In order to identify the mechanisms involved, the production of TGF-β1, CCL3, IFN-γ, TNF-α, IL-10, VEGF, IL-6, IL-1β, IL-17 and IL-2 in the lung homogenate was evaluated. Our findings revealed that infected mice treated with PTX/ITC s... (Complete abstract click electronic access below) / Mestre

Cystic fibrosis (CF) is an inborn, hereditary disease, due to mutations in the gene for a cAMP-activated chloride (Cl-) channel, the cystic fibrosis transmembrane conductance regulator (CFTR). As a result of impaired ion and water transport,the airway mucus is abnormally viscous, which leads to bacterial colonization.Recurrent infections and inflammation result in obstructive pulmonary disease.Similar changes in the pancreas lead to pancreatic insufficiency.Several compounds have been tested to improve transepithelial ion transport in CF patients, either via activation of the mutant CFTR, or via stimulation of alternative chloride channels. The main purpose of this thesis was to find substances that might correct the defective ion transport in epithelial cells in CFand could be useful for the pharmacological treatment of CF patients. Long-term treatment with the macrolide antibiotic azithromycin (AZM)improved clinical parameters and lung function in CF patients and increased Cl- transport in CF bronchial epithelial cells (CFBE) (Paper I); although mRNA expression of the CFTR gene remained unchanged.In contrast, pre-exposure to the mucolytic antioxidant N-acetylcysteine (NAC) increased CFTR protein expression and was associated with increased Cl- efflux from CFBE cells (Paper II). Clinical trials of this substance might be warranted. Duramycin has been the subject of clinical trials that finished in June2009. Up till now, no results from this study are available. The effect of this substance on Cl- efflux from three CF and three non-CF cell lines (Paper III) was disappointing. An effect was found only in CFBE cells, the effect was minimal, occurred in a narrow concentration range, and was not associated with an increase in the intracellular calcium concentration [Ca2+]i. The fact that NO-donors stimulated Cl- efflux from CFBE cells (but did notchange [Ca2+]i) after several hours of preincubation suggests that these substances may be a potentially interesting group of compounds for the treatment of CF (Paper IV). A model for the effect of NO-donors on Cl- efflux is presented. / Cystisk fibros (CF) är en medfödd, ärftlig, sjukdom, som förorsakas av en mutation i en gen som innehåller koden för en kloridkanal som aktiveras av cykliskt AMP (cystic fibrosis transmembrane conductance regulator, CFTR). Som en följd av otillräcklig transport av joner och vatten är slemmet i luftvägarna onormalt segt, vilket leder till att det koloniseras av bakterier. Upprepade infektioneroch inflammation av luftvägarna leder slutligen till obstruktiv lungsjukdom.Liknande förändringar i bukspottkörteln leder till att också detta organ inte fungerar. Flera kemiska ämnen har testats för sin förmåga att förbättra jontransporten över epitelet hos CF-patienter. Detta skulle kunna göras antingen genom aktivering av det muterade CFTR-proteinet, eller genom stimulering av alternativa kloridkanaler. Huvudsyftet med den forskning som beskrivs i denna avhandling var att hitta kemiska substanser som skulle kunna korrigera den defekta jontransporten i epitelceller hos CF-patienter, och därför vara nyttiga för behandlingen av patienterna. Behandling under längre tid med azithromycin (AZM), ett makrolidantibiotikum,förbättrade CF-patienternas kliniska status och lungfunktion,samt ökade kloridutflödet från CF bronkialepitelceller (CFBE-celler) (Arbete I).Däremot ändrades inte uttrycket av mRNA för CFTR-genen. I kontrast till detta ökade uttrycket av CFTR-proteinet om CFBE-cellerna utsattes för den slemlösande anti-oxidanten N-acetylcystein (NAC), vilket ledde till ökat kloridutflöde från denna cellinje (Arbete II). Det vore rimligt att utföra kliniska prövningar av detta ämne. Duramycin har testats i kliniska prov som slutade i juni 2009, men några resultatfrån dessa prov har inte offentliggjorts än. Effekten av detta ämne på kloridutflödet från tre CF-cellinjer och tre icke-CF cellinjer (Arbete III) var en besvikelse. Duramycin hade endast effekt på CFBE-celler, effekten var mycket liten, förekom endast i ett litet koncentrationsområde av duramycin, och var inte kopplad till en ökning av den intracellulära kalciumkoncentrationen [Ca2+]i. Att ämnen som avger kväveoxid (NO) stimulerade kloridutflödet från CFceller (men inte påverkade [Ca2+]i) efter några timmar, visar att denna grupp av ämnen kan vara potentiellt intressant för behandlingen av CF (arbete IV). En modell för effekten av NO på kloridtransporten i CF-celler presenteras.

Background: Clinical breakpoints and epidemiological cut-off values for N. gonorrhoeae azithromycin antimicrobial susceptibility testing have not been established. This study utilized existing minimum inhibitory concentration (MIC) data from CDC’s Gonococcal Isolate Surveillance Project (GISP) to establish epidemiological cut-off values for azithromycin and N. gonorrhoeae as determined by agar dilution. Methods: MIC distributions for the pooled dataset and each data year (2005-2012) were constructed. Epidemiological cut-off values were calculated using two methods. Method 1 considers the wild-type MIC distribution, the modal MIC for the distribution, and the inherent variability of the test (±1 twofold-dilution). Method 2 defines the epidemiological cut-off value as two twofold-dilutions higher than the MIC50. Results: Taking into consideration the wild-type MIC distributions and the inherent variability of the test, the epidemiological cut-off value chosen for the pooled dataset and each data year using Method 1 was ≤1.0 µg/mL. The MIC50 for the pooled dataset and each data year was 0.25 µg/mL. Two twofold-dilutions higher than the MIC50 (0.25 µg/mL) for the pooled dataset and each data year was 1.0 µg/mL. Discussion: The epidemiological cut-off values chosen using Methods 1 and 2 (≤1.0 µg/mL) were identical for the pooled dataset and each data year, indicating the epidemiological cut-off value has not changed from 2005-2012. The epidemiological cut-off value for N. gonorrhoeae azithromycin agar dilution antimicrobial susceptibility testing established during this study can be used to help set clinical breakpoints and identify isolates with reduced susceptibility to azithromycin.

Azithromycin improves clinical outcomes in patients with cystic fibrosis (CF), specifically in patients infected with Pseudomonas aeruginosa. Azithromycin shifts macrophage programming away from a pro-inflammatory classical (M1) phenotype, and towards an anti-inflammatory alternative (M2) phenotype; however, little is known about this mechanism, nor of its impact upon immune response to pulmonary infection. We set out to determine the mechanism by which azithromycin is able to alter macrophage phenotype, and assess the effect of azithromycin induced macrophage polarization on inflammation during pulmonary infections. Utilizing macrophage cell culture, we found that azithromycin increased IKKβ, a signaling molecule in the NFκB pathway, which likely is altering macrophage programming. Using a Pseudomonas infection model in mice that lack physiologic alternative macrophage activation, we showed that azithromycin’s ability to alter macrophage function and decrease lung damage was independent of interleukin control of macrophage programming. Azithromycin increased fibrotic protein production both in vivo and in vitro, but blunted immune-driven fibrotic damage. We extended our study to patients with CF, describing gene expression in macrophages isolated from sputum samples. We found markers consistent with a shift toward M2 polarization in these patients. These data suggest potential mechanisms by which azithromycin benefits patients with CF.

The Plasmodium falciparum parasite, carried by Anopheles mosquitoes, is currently a global problem due to the rising incidence of resistance of the parasite to available antimalaria drugs. Resistance and difficult treatment groups, including pregnant woman and young children, are pressing for the development of new, safe and effective prophylactic and treatment antimalarials. Because of the extensive process of developing new drugs, researchers and health care professionals have turned to combination therapy where a fast acting antimalarial is combined with slower acting drugs, such as antibiotics. The macrolide antibiotics, erytbromycin and azithromycin, have been studied to a limited extent for their potential antimalarial effect. Certain advantages, such as their safety profile (especially that of azithromycin) in pregnancy and administration to young children, motivates continual research into the advancement of the effect these drugs exude on malaria. Drug delivery systems contribute to the efficacy of medicines, conquering several difficulties of treatment with oral medication. Pheroid™ technology is a patented drug delivery system, mainly consisting of plant and essential fatty acids, and has been demonstrated to entrap, carry and deliver pharmacologically active compounds and other useful molecules. This study compared the in vitro effects of the macrolide antibiotics on the growth of a chloroquine-resistant strain (RSA 11) of Plasmodium falciparum to the effects of the macrolides entrapped in Pheroid™ vesicles on the same strain over and extended observation period of 144 hours. ELISA assays were conducted by analysing the HRP II (histidine-rich protein) levels on a pre-coated microtitre plate. The effects of the type of formulation, concentration and time were compared. The in vitro difference between erythromycin alone and entrapped in Pheroid™ vesicles were found to be statistically significant (p = 0.000000) while the effects of both formulations did not seem to be concentration dependant (p = 0.628424). Prolonged exposure was also statistically meaningful (p = 0.008268), though it seems that exposure need not exceed 96 hours. The type of formulation, in the case of azithromycin (azithromycin alone vs. azitbromycin entrapped in Pheroid™ vesicles), proved statistically significant (P = 0.002572), while neither formulation seemed concentration dependant (P = 0.427731). Prolonged exposure was found to be statistically insignificant for azithromycin (P = 0.221941). / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2010.

The Plasmodium falciparum parasite, carried by Anopheles mosquitoes, is currently a global problem due to the rising incidence of resistance of the parasite to available antimalaria drugs. Resistance and difficult treatment groups, including pregnant woman and young children, are pressing for the development of new, safe and effective prophylactic and treatment antimalarials. Because of the extensive process of developing new drugs, researchers and health care professionals have turned to combination therapy where a fast acting antimalarial is combined with slower acting drugs, such as antibiotics. The macrolide antibiotics, erytbromycin and azithromycin, have been studied to a limited extent for their potential antimalarial effect. Certain advantages, such as their safety profile (especially that of azithromycin) in pregnancy and administration to young children, motivates continual research into the advancement of the effect these drugs exude on malaria. Drug delivery systems contribute to the efficacy of medicines, conquering several difficulties of treatment with oral medication. Pheroid™ technology is a patented drug delivery system, mainly consisting of plant and essential fatty acids, and has been demonstrated to entrap, carry and deliver pharmacologically active compounds and other useful molecules. This study compared the in vitro effects of the macrolide antibiotics on the growth of a chloroquine-resistant strain (RSA 11) of Plasmodium falciparum to the effects of the macrolides entrapped in Pheroid™ vesicles on the same strain over and extended observation period of 144 hours. ELISA assays were conducted by analysing the HRP II (histidine-rich protein) levels on a pre-coated microtitre plate. The effects of the type of formulation, concentration and time were compared. The in vitro difference between erythromycin alone and entrapped in Pheroid™ vesicles were found to be statistically significant (p = 0.000000) while the effects of both formulations did not seem to be concentration dependant (p = 0.628424). Prolonged exposure was also statistically meaningful (p = 0.008268), though it seems that exposure need not exceed 96 hours. The type of formulation, in the case of azithromycin (azithromycin alone vs. azitbromycin entrapped in Pheroid™ vesicles), proved statistically significant (P = 0.002572), while neither formulation seemed concentration dependant (P = 0.427731). Prolonged exposure was found to be statistically insignificant for azithromycin (P = 0.221941). / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2010.

Azithromycin (AZ) is a macrolide antibiotic derivate from erytromycin. AZ has a broad spectrum of activity against common gram-negative pathogens and has been used for the treatment of respiratory tract infection, skin infections and sexually transmitted diseases. In the Brazilian market AZ is available as tablets, compounded capsules and powder for oral suspension. The official methods for the assay of AZ in bulk form; capsules and powder for oral suspension are high performance liquid chromatography or microbiological diffusion assay. No monographs are reported in the pharmacopoeias for AZ evaluation in tablets. In this work two spectrophotometric methods were developed and validated for AZ analysis in tablets and powder for oral suspension. The first method was based on the reaction of AZ with concentrated sulfuric acid (98 % w/v), with detection at 226 nm. The other method was based in the charge-transfer reaction of the drug with s-acceptor iodine, with detection at 363 nm. Both methods showed good linearity (r>0.99), precision (CV<5%) and accuracy (>99%). The results obtained with the proposed methods were in good agreement with those obtained by microbiological diffusion agar method. The optimization of dissolution test conditions for in vitro quality control of AZ in tablets was also studied. The use of 900 mL of 0.1N HCl at 37.0 ± 0.5 ºC, paddle as apparatus, at a stirring rate of 50 rpm, provided satisfactory results for tested products. The percent dissolution of AZ in the established condition was more than 90% in 45 minutes. The validated spectrophotometric method used to evaluate the dissolution testing showed to be specific (with no interference of the placebo or tablets in the quantification of AZ), linear (r>0.99), precise (RSD<5%) and accurate (>97%). The drug showed satisfactory stability in the selected dissolution medium / Azitromicina (AZ) é um antibiótico macrolídeo derivado da eritromicina. AZ apresenta um grande espectro de atividade contra patógenos Gram negativos comuns e tem sido utilizada para o tratamento de infecções do trato respiratório, infecções da pele e doenças sexualmente transmissíveis. No mercado brasileiro a AZ está disponível na forma de comprimidos, cápsulas manipuladas e pó para suspensão oral. Os métodos oficiais para o doseamento de AZ como matéria-prima; cápsulas e pó para a suspensão oral são a cromatografia líquida de alta eficiência ou o método microbiológico de difusão em ágar. Não existem monografias em farmacopéias para avaliação de AZ em comprimidos. Neste trabalho dois métodos espectrofotométricos foram desenvolvidos e validados para a análise de AZ em comprimidos e em pó para suspensão oral. O primeiro método foi baseado na reação de AZ com ácido sulfúrico concentrado (98%, p/v), com detecção em 226 nm. O outro método foi baseado na reação de complexo de transferência de carga do fármaco com o s-aceptor iodo, com detecção em 363 nm. Ambos os métodos mostraram linearidade (r>0,99), precisão (CV<5%) e exatidão (>99%). Os resultados obtidos com os métodos propostos estavam de acordo com aqueles obtidos com o método microbiológico de difusão em ágar. A otimização das condições do teste de dissolução para o controle de qualidade in vitro de AZ em comprimidos foi também estudado. O uso de 900 mL de HCl 0,1N, a 37 ºC ± 0,5 ºC, aparato pá, a um a velocidade de 50 rpm, demonstrou resultados satisfatórios para produtos testados. A percentagem de dissolução de AZ na condição estabelecida foi superior a 90% em 45 minutos. O método espectrofotométrico validado para avaliar a dissolução mostrou-se específico, sem interferência do placebo dos comprimidos, na quantificação de AZ, linear (r>0.99), preciso (CV< 5%) e exato (>97%). O fármaco mostrou estabilidade satisfatória no meio de dissolução selecionado

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2018-05-21T14:55:49Z No. of bitstreams: 1 PDF - Mysrayn Yargo de Freitas Araújo Reis.pdf: 25441289 bytes, checksum: 518beaa4a3b4581b9a50e243986be6b6 (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2018-05-23T16:59:36Z (GMT) No. of bitstreams: 1 PDF - Mysrayn Yargo de Freitas Araújo Reis.pdf: 25441289 bytes, checksum: 518beaa4a3b4581b9a50e243986be6b6 (MD5) / Made available in DSpace on 2018-05-23T16:59:36Z (GMT). No. of bitstreams: 1 PDF - Mysrayn Yargo de Freitas Araújo Reis.pdf: 25441289 bytes, checksum: 518beaa4a3b4581b9a50e243986be6b6 (MD5) Previous issue date: 2017-03-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Bacterial Skin Infections (BSIs) includes a varied group of diseases caused by the invasion of bacteria into the epidermidis and adjacent soft tissues. Although the skin can host bacteria belonging to the normal cutaneous microbiota, such as, several speciessuch as, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa can be considered pathogens for several IBPs. Among the various antibiotic groups used in treatment of BSIs, azithromycin is mentioned in the literature. Azithromycin (AZ), is a semi-synthetic macrolide antibiotic of the azalide class and is a promising candidate for the treatment of BSIs due to their capacity of penetration in the tissues and wide spectrum of action for several bacterial species. The incorporation of AZ into new drugs delivery systems such as microemulsion (ME) can ensure release control, lower toxicity, lower doses, decrease in plasma peaks, protection of drug degradation or inactivation. The present work aims to encapsulate AZ into microemulsion systems, perform the characterization and confirm its antibacterial activity. From the pseudoternary phase diagram were developed and selected one type of O/W formulations composed of deionized water, isopropyl myristate, LAS , and Brij 52 for the encapsulation of AZ. Both formulations in the absence (MEB) and presence of drug (MEAZ) were presented as clear and stable systems. For MEAZ, the pH value (6.75) indicated an excellent compatibility for dermal administration. In addition, its high conductivity (630.1), allied with the differential scanning calorimetry (DSC) studies, suggested an O/A type system. The refractive index revealed the isotropic character, confirmed by the technique of polarized light microscopy. Transmission electron microscopy (MET) indicated the nanometric droplet size of the systems and the dynamic light scattering (DLS) technique was able to measure a diameter of 18.72 and 20.89 nm for MEB and MEAZ, respectively. From the rheological analysis we can attest to the Newtonian type behavior for both formulations. Finally, the antimicrobial activity of MEAZ against the strains of S.aureus, S.epidermidis and P.aeruginosa was confirmed. Studies necessary to trace the release profile and determination of minimal inhibitory concentration will be conducted so that the developed ME may represent a new safe and effective alternative in the complementary treatment of PPIs. / As infecções bacterianas da pele (IBPs) engloba um variado grupo de doenças causadas pela invasão de bactérias na epiderme e tecidos moles adjacentes. Embora a pele possa abrigar bactérias que pertencem a microbiota cutânea normal, muitas espécies tais como, Staphylococcus aureus, Staphylococcus epidermidis e Pseudomonas aeruginosa podem ser consideradas agentes patógenos para várias IBPs. Dentre os vários grupos de antibióticos utilizados no tratamento das IBPs, pode ser citada a azitromicina (AZ). A AZ é um antibiótico macrolídeo semi-sintético da classe dos azalídeos, apresenta-se como um promissor candidato no tratamento das IBPs uma vez que possui a capacidade de penetração nos tecidos e largo espectro de ação para diversas espécies bacterianas. A incorporação de AZ em novos sistemas de liberação de fármacos visando a sua administração por via tópica, como as microemulsões (ME), pode garantir o controle da liberação, menor toxicidade, menor número de doses, diminuição dos picos plasmáticos, a proteção da degradação e/ou inativação do fármaco. O presente trabalho teve como objetivo incorporar a AZ em uma ME, realizar a sua caracterização e confirmar a sua atividade antibacteriana. A partir do diagrama de fases pseudoternário foi desenvolvida e selecionada uma microemulsão do tipo O/A compostas por água deionizada (fase aquosa), miristato de isopropila (fase oleosa), LAS e Brij 52 (tensoativos na proporção 9:1) para a incorporação de AZ. Ambas as formulações na ausência (MEB) e presença de fármaco (MEAZ) se apresentaram como sistemas límpidos e estáveis. Para a MEAZ, o valor de pH (6,75) indicou uma ótima compatibilidade para administração cutânea. Além disso, a sua elevada condutividade (630, 1µScm^-1), aliada aos estudos de calorimetria exploratória diferencial (DSC) sugeriram um sistema do tipo O/A. O índice de refração revelou o caráter isotrópico, confirmada pela técnica de microscopia de luz polarizada. A microscopia eletrônica de transmissão (MET) indicou o tamanho de gotículas nanométrico dos sistemas e a técnica de espalhamento de luz dinâmica (DLS) foi capaz de medir um diâmetro de 18,72 e 20,89 nm, para a MEB e MEAZ, respectivamente. A partir de análises reológicas pode-se atestar o comportamento do tipo newtoniano para ambas as formulações. Por último, foi atestada a atividade antimicrobiana da MEAZ contra as cepas de S. aureus, S.epidermidis e P. aeruginosa. Estudos necessários para traçar o perfil de liberação e determinação da concentração inibitória mínima serão conduzidos para que a ME desenvolvida possa representar uma nova alternativa segura e eficaz no tratamento complementar das IBPs.

Several chlamydial antigens have been detected in the infected epithelial cell cytosol and on the host cell surface prior to their presumed natural release at the end of the 72-96 h developmental cycle. These extra-inclusion antigens are proposed to influence vital host cell functions, antigen trafficking and presentation and, ultimately, contribute to a prolonged inflammatory response. To begin to dissect the mechanisms for escape of these antigens from the chlamydial inclusion, which are enhanced on exposure to antibiotics, polarized endometrial epithelial cells (HEC-1B) were infected with Chlamydia trachomatis serovar E for 36 h or 48 h. Infected cells were then exposed to chemotactic human polymorphonuclear neutrophils not loaded or pre-loaded in vitro with the antibiotic azithromycin. Viewed by electron microscopy, the azithromycin-mediated killing of chlamydiae involved an increase in chlamydial outer membrane blebbing followed by the appearance of the blebs in larger vesicles (i) everting from but still associated with the inclusion as well as (ii) external to the inclusion. Evidence that the vesicles originated from the chlamydial inclusion membrane was shown by immuno-localization of inclusion membrane proteins A, F, and G on the vesicular membranes. Chlamydial heat shock protein 60 (chsp60) copies 2 and 3, but not copy 1, were released from RB and incorporated into the everted inclusion membrane vesicles and delivered to the infected cell surface. These data represent direct evidence for one mechanism of early antigen delivery, albeit membrane-bound, beyond the confines of the chlamydial inclusion.