Introdução: Há crescentes evidências que sinalizam o papel do estresse crônico como desencadeante e mesmo perpetuador de crises asmáticas, bem como a importância da produção de muco e do transporte mucociliar na fisiopatologia da asma brônquica. Objetivos: Deste modo, consideramos relevante avaliar em cobaias com inflamação alérgica crônica pulmonar como o estresse físico repetido, induzido pela natação forçada, modula as respostas de transporte mucociliar, as propriedades do muco, o infiltrado eosinofílico e a expressão de IL-13 no epitélio e nas células inflamatórias na parede das vias aéreas. Métodos: Os animais receberam inalações duas vezes por semana durante quatro semanas com doses crescentes de ovoalbumina (grupo OVA e OVA-NAT) ou solução fisiológica (grupo SAL e SAL-NAT). Após 24 horas da quarta inalação os animais dos grupos denominados SAL-NAT e OVA-NAT foram submetidos ao protocolo de natação forçada por dez dias com intervalo de dois dias, para a indução do estresse. Após 72 horas da última inalação, os animais foram anestesiados, sendo testadas: a velocidade de transporte mucociliar (VTM), a freqüência de batimento ciliar (FBC), e a diferença de potencial transepitelial (PD). Após estas medidas foram coletadas amostras de muco para avaliação do ângulo de contato (AC) e da transportabilidade pela tosse (TT). Foi coletada uma amostra de sangue para a dosagem de cortisol sérico. Os pulmões foram retirados, fixados e submetidos à coloração de LUNA, para identificação de eosinófilos, e à técnica imunohistoquímica para detecção da expressão de IL-13 no epitélio brônquico e nas células inflamatórias presentes na parede das vias aéreas. As adrenais também foram retiradas, imediatamente pesadas e submetidas à coloração de hematoxilina e eosina para avaliação histopatológica, utilizando a morfometria para determinação das áreas de cada uma de suas camadas. Resultados: Os Resumo animais do grupo OVA apresentaram uma redução da VTM, do PD, da TT e aumento do AC, da área de epitélio de vias aéreas, de muco ácido, do número de eosinófilos e da expressão de IL-13, no epitélio brônquico e nas células inflamatórias presentes ao redor das vias aéreas, comparativamente aos controles (p<0,05 para todas as comparações). Houve redução da área total da adrenal e de cada uma de suas camadas comparativamente aos controles (p<0,05 para todas as comparações). Os animais submetidos ao protocolo de estresse (grupos SAL-NAT e OVA-NAT) apresentaram aumento nos níveis de cortisol sérico (p<0,05), no peso das adrenais comparativamente aos grupos e OVA (p<0,05). Os animais submetidos ao protocolo de estresse e expostos à ovoalbumina (grupo OVA-NAT) mostraram redução na VTM, aumento no AC, na área de muco ácido do epitélio brônquico e em todas as camadas das adrenais, comparativamente aos resultados obtidos nos animais do grupo OVA (p<0,05 para todas as comparações). Conclusão: O processo inflamatório crônico pulmonar altera o transporte mucociliar e as propriedades reológicas do muco, o que se associou ao recrutamento eosinofílico, aumento da expressão de IL-13 no epitélio brônquico e nas células inflamatórias presentes na parede brônquica. Além disto, observamos a redução de todas as camadas da adrenal. Neste modelo experimental, o estresse físico repetido reduz o transporte mucociliar devido a alterações das propriedades do muco, particularmente potencializando o aumento de muco ácido e de sua hidrofobicidade e adesividade. Estes efeitos parecem estar relacionados à ativação adrenal, mas independentes do recrutamento eosinofílico e da resposta Th2 mediada pela IL-13 / Background: It has increasing evidence linking the role of stress in the onset of asthma exacerbation and in the maintenance of asthmatic crises, as well as the importance of mucus production and the mucociliary clearance in the asthma physiopathology. Objectives: So, we consider relevant to evaluate in guinea pigs with pulmonary chronic allergic inflammation how the induced repeated physical stress, caused by forced swimming, modulates the mucociliary clearance, the mucus properties, the eosinophilic infiltration and the IL-13 expression on bronchial epithelial cells in the airway wall. Methods: The animals had received inhalations two times per week during four weeks with increasing doses of ovalbumin (OVA and OVA-S groups) or saline solution (SAL and SAL-S groups). After twenty four hours of the 4th inhalation, the animals (named as SAL-S and OVA-S groups) had been submitted to the protocol of forced swimming, per ten days with an interval of two days, for the stress induction. After 72 hours of the last inhalation, the animals were anaesthetized and the tracheal mucus clearance (TMC), the ciliary beat frequency (CBF), and the difference of transepithelial potential difference (PD) were measured. After these measurements had been done mucus samples were collected for evaluation of the contact angle (CA) and cough transportability (CT). To serum cortisol dosage a little amount of blood was collected. The lungs were dissected, fixed and submitted to histological techniques, like LUNA stain for identification of eosinophils and the IL-13 immunohistochemistry technique for its detection in the bronchial epithelium and in the inflammatory cells present in the airways walls. The adrenal glands were excised, immediately weighed and submitted to haematoxylin and eosin stain for histopathological evaluation using the morphometry to determine the areas of each of their layers. Results: The animals of OVA Summary group presented a reduction of the TMC, the PD, and the CT. On the other hand they presented an increase of the CA, of the airways epithelial area, of the acid mucus, of the eosinophils number as well as on the IL-13 expression in the airways epithelium and on the inflammatory cells around the airways, compared to the controls (p<0.05 for all comparisons). There was a reduction of the adrenal gland total area and on each one of its layers compared to the controls (p<0.05 for all comparisons). The animals submitted to the forced swim stress protocol (SAL-S and OVA-S groups) showed an increase on serum cortisol levels, on adrenal gland weight compared to OVA group (p<0.05). The animals submitted to the forced swim stress protocol and to the sensitization protocol with ovalbumin (OVA-S) showed a reduction in the TMC and an increase on the CA, bronchial epithelial acid mucus area and in all adrenal gland layers compared to the results obtained in the OVA group animals (p<0.05 for all comparisons). Conclusions: The chronic inflammatory pulmonary process alters the mucociliary transport and mucus rheological properties which was associated to eosinophilic recruitment, increases in the IL-13 expression on bronchial epithelial cells and inflammatory cells in airway walls. In addition, we observed a reduction in all adrenal zones. In this experimental model the repeated physical stress reduces the mucociliary clearance due to the mucus rheological properties alterations, particularly increasing the amount of acid mucus and its wettability and adhesivity. These effects seem to be related to the adrenal activation but independently of the eosinophilic recruitment and of Th2 responses mediated by IL-13

Base teórica: Muitos pacientes internados em Unidade de Terapia Intensiva (UTI) necessitam da instituição da ventilação mecânica invasiva (VM). Porém esta pode trazer efeitos deletérios como: alteração na higiene brônquica e pneumonia associada à ventilação mecânica (PAV). Devido a isso, faz-se necessária avaliação de técnicas de higiene brônquicas eficazes e o menos deletérias possíveis. Objetivos: Comparar a eficácia das técnicas em estudo: vibrocompressão (VB) (G1), hiperinsuflação com o ventilador mecânico (HMV) (G2) VB + HMV (G3) na quantidade de secreção aspirada (SEC), no tempo de VM, na incidência de PAV, na reintubação orotraqueal (Re-IoT) e na mortalidade de pacientes em VM. Método: Ensaio clínico randomizado, realizado no Centro de Terapia Intensiva do Hospital de Clínicas de Porto Alegre (HCPA). Foram incluídos no estudo 93 pacientes (29 G1, 32 G2 e 32 G3) em VM por mais de 24 horas. Foram aplicadas as seguintes técnicas: aspiração isolada (ASP), VB, HMV e VB + HMV. Foram medidas as seguintes variáveis: frequência cardíaca (FC), frequência respiratória (FR), pressão arterial média (PAM), saturação arterial periférica de oxigênio (SpO2), pressão inspiratória de pico (PIP), volume corrente (VC), complacência dinâmica (Cdyn), peso da SEC; tempo de VM, Re-IoT, incidência de PAV e mortalidade na VM. Conclusão: O grupo 3 foi o único que apresentou aumento significativo da SEC, quando comparado à ASP (0,7 g (0,1-2,5) vs 0,2 g (0,0-0,6) – p = 0,006). Em comparação com os demais grupos, o grupo 2 apresentou aumento significativo na incidência de PAV (22% - p = 0,003) e Re-IoT (21,9% - p = 0,048), demonstrando assim, na amostra estudada, que VB + HMV é mais eficaz quanto à quantidade de SEC e exerce efeito protetor, juntamente com a VB, na incidência de PAV e Re-IoT. / Background: Many patients admitted to the Intensive Care Unit (ICU) require the institution of invasive Mechanical Ventilation (MV). However, this can bring harmful effects such as changes in mucociliary transport and cough capacity, leading to bronchial obstruction and Ventilator Associated-Pneumonia (VAP). Objective: To compare the efficacy of the techniques: Vibrocompression (G1), Hyperinflation with Mechanical Ventilation (G2) Vibrocompression + Hyperinflation with Mechanical Ventilation (G3) in the amount of Aspirated Secretions (AS), MV time, the incidence of VAP, Re-intubation tracheal (Re-IoT) and mortality of patients on MV. Method: Randomized clinical trial, conducted at the Intensive Care Unit, of the Hospital de Clinicas de Porto Alegre (HCPA). 93 patients were included in the study (29 G1, 32 G2 and 32 G3) in mechanical ventilation for more than 24 hours. The following techniques were applied: Isolated Aspiration (ASP), Vibrocompression (VB), Hyperinflation with Mechanical Ventilation (HMV) Vibrocompression + Hyperinflation with Mechanical Ventilation (HMV + VB). The following variables were measured: Heart Rate (HR), Respiratory Rate (RR), Mean Arterial Pressure (MAP), Peripheral Arterial Oxygen Saturation (SpO2); Peak Inspiratory Pressure (PIP), Tidal Volume (TV); Dynamic Compliance (Cdyn); weight of the AS; VM time; Re-IoT; VAP incidence and mortality in the VM. Conclusion: The HMV + VB (G3) was the only group that showed significant increase in the AS (0.7 g (0.1-2.5) vs 0.2 g (0.0-0.6) – p = 0.006) when compared to ASP. Compared with the other groups G2 showed a significant increase in the incidence of VAP (22% - p = 0.003) and Re-IoT (21.9% - p = 0.048). Thus demonstrating in the sample, which HMV + VB is more effective as the amount of secretion aspirated and has a protective effect, along with the VB, the incidence of VAP and Re-IoT.

O objetivo do presente trabalho foi propiciar o acesso in vivo ao epitélio respiratório e estudar a frequência de batimento ciliar (FBC) e a diferença de potencial transepitelial (DP) em um modelo murino de doença alérgica das vias aéreas induzida por ovoalbumina. Camundongos Swiss foram sensibilizados com ovoalbumina (OVA) através de duas injeções intraperitoneais de alérgico com o adjuvante hidróxido de alumínio (dias 0 e 14) e quatro inalações de OA 1% (dias 22, 24, 26 e 28). O grupo controle (S) foi tratado com salina 0,9 % seguindo o mesmo protocolo. Após 48h da última inalação, os camundongos foram anestesiados, a traquéia foi exposta longitudinalmente (1x4 mm) e o epitélio pode ser visualizado. A FBC foi mensurada pela técnica estroboscópica antes (basal) e logo após a administração inalatória das drogas (salbutamol e brometo de ipratrópio). A DP foi mensurada nos grupos S e OVA. Foram avaliados o lavado broncoalveolar e o remodelamento do epitélio da cavidade nasal, traquéia e vias aéreas distais. Nenhuma diferença foi encontrada na FBC basal entre os grupos (OVA e S), no entanto o grupo OVA mostrou uma DP basal significativamente menor. A inalação de salbutamol (3.5.10-3M ou 3.5.10-4M) elevou a FBC nos grupos estudados (p<0,05). O brometo de ipratrópio (10- 4M e 6.10-4M) não influenciou a FBC basal. Nossos resultados mostraram que é possível avaliar a FBC e a DP in vivo em um modelo murino de doença pulmonar alérgica crônica, e indicam que o processo inflamatório não afeta a FBC, mas contribui para o aumento de muco nas vias aéreas com conseqüências deletérias ao transporte mucociliar facilitando a retenção / The aim of the present work was to propitiate the in vivo assessment of the respiratory epithelium. The effects of salbutamol and ipratropium bromide on ciliary beat frequency (CBF) in a murine model of allergic airway disease were addressed. Transepithelial electric potential difference (PD) was also measured in order to verify the integrity of the epithelial barrier. Mice were sensitized with ovalbumin (OVA) by two intraperitoneal injections of allergen (days 0 and 14) and four inhalations of OVA 1% (days 22, 24, 26 and 28). The control group was treated with saline following the same procedures. After 48 hs of the last inhalation, mice were anesthetized, trachea was opened longitudinally (1 x 4 mm) and the ciliated epithelium could be visualized. CBF was measured by a modification on the videoscopic technique. We measured the CBF before and just after the administration of aerosolized substances. The PD was also measured on groups OVA and S. Additionally, the eosinophil cell count was measured on broncoalveolar lavage (BAL) in order to access the magnitude of airway inflammation. No difference on baseline CBF was noticed between groups (OVA and S), however the OVA group had a significantly lower PD. The administration of aerosolized capsaicin (3.10-9M) and salbutamol (3.5.10-3M or 3.5.10-4M) increased CBF in all groups studied. Ipratropium bromide (10-4M and 6.10- 4M) did not influence the CBF. The eosinophil cell count in broncoalveolar lavage was higher in OVA group compared to S group. CBF and PD results indicate that the inflammatory process does not affect the ciliary beat frequency but augments the amount of mucus in the airway, with deleterious consequences to the mucociliary transport facilitating mucus retention. Our results demonstrated for the first time the possibiliy of studying airway epithelium in an in vivo murine model of allergic airway disease

Made available in DSpace on 2015-04-22T22:10:32Z (GMT). No. of bitstreams: 1 Enily Vieira.pdf: 1694755 bytes, checksum: 6d8dfa6ad931b649316127bb22700791 (MD5) Previous issue date: 2011-12-13 / The airport infrastructure and services of customs clearance of imported cargo can be considered a competitive advantage of companies in the Industrial Pole of Manaus (PIM). Reducing the time of cargo clearance would be just one advantage. The study aims to propose a management model agile and flexible in receiving air cargo imported inputs so that it satisfactorily meets the agile supply chain, which has the ability to respond to change, diversity, the unpredictable demands of customers, and which reduces the risk of pipeline breaks for companies in the PIM. This work, which culminates with a case study not only addresses the manufacturers of high-tech and innovative products that are of high value added, but all the actors (INFRAERO, RFB, ANVISA, MAP, broker, importing companies) involved import cargo, since his arrival in TECA to the delivery of cargo to the importer. The research call attention to the importance of infrastructure and services in the fast release of imported cargo through a management model able to include in the PIM requirements of external competitiveness of supply chains, using appropriate strategies such as model uncertainty supply chain (MICS). As a result, obtained presents an alternative model identified as the Special Customs Regime of Industrial Warehouse on Computerized Control (Ricof), considered an instrument of attraction and facilitator of global business development of products considered as part of the chain of uncertainty, so that gain is observed not only the logistical and tax, knowing that in Brazil there are no special schemes and atypical, as the companies of the Industrial Pole of Manaus (PIM). / A infraestrutura aeroportuária e os serviços de desembaraço aduaneiro de cargas importadas podem ser considerados uma vantagem competitiva das empresas no Pólo Industrial de Manaus (PIM). A redução do tempo de liberação de cargas seria apenas uma das vantagens. O estudo tem como objetivo propor um modelo de gerenciamento ágil e flexível na recepção de cargas aéreas de insumos importados de modo que este atenda satisfatoriamente a Cadeia de Suprimentos Ágil, que tem a capacidade de responder as mudanças, diversidades, demandas imprevisíveis dos clientes, e que reduz os riscos de interrupções dos fornecimentos para as empresas do PIM. Este trabalho, que culmina com um estudo de caso, não somente aborda os fabricantes de produtos high-tech e inovadores, que são de alto valor agregado, mas todos os atores (INFRAERO, RFB, ANVISA, MAPA, DESPACHANTE, Empresas Importadoras) envolvidos na importação da carga, desde sua chegada no TECA até a entrega da carga para o importador. A pesquisa dá visibilidade à importância da infraestrutura e rapidez nos serviços de liberação de cargas importadas, através de um modelo de gerenciamento capaz de incluir o PIM nas exigências da competitividade externa entre as cadeias de suprimento, fazendo uso de estratégias adequadas como o modelo de incerteza da cadeia de suprimentos (MICS). Como resultado obtido é apresentado um modelo identificado como alternativa ao Regime Aduaneiro Especial de Entreposto Industrial sobre Controle Informatizado (RICOF), considerado instrumento de atração e facilitador de desenvolvimento das empresas globais de produtos considerados como integrantes da cadeia de incerteza, de modo que seja observado o ganho logístico e não apenas o tributário, sabendo que no Brasil não se inclui os regimes especiais e atípicos, como as empresas do Pólo Industrial de Manaus (PIM).

O tabagismo está fortemente associado ao desenvolvimento da Doença Pulmonar Obstrutiva Crônica, porém poucos estudos que relatam alterações funcionais ou inflamatórias nas vias aéreas superiores em jovens são encontrados na literatura. Nosso objetivo foi investigar as alterações funcionais e inflamatórias nas vias aéreas superiores e inferiores em jovens tabagistas com idade igual ou inferior a 35 anos. Setenta e dois indivíduos participaram do estudo: 32 jovens não tabagistas (JNT) saudáveis (21±4 anos, 29 homens) e 40 jovens tabagistas subdivididos de acordo com a carga tabágica: menor que 2,5 anos-maço (< 2,5) (19±2 anos, 20 homens) e igual ou maior que 2,5 anos-maço (>= 2,5) (24 ± 5 anos, 17 homens e três mulheres). Foram avaliados os dados demográficos, os dados clínicos, os sintomas de desconforto das vias aéreas por meio do questionário SNOT-20, os volumes e capacidades pulmonares por meio do teste de função pulmonar, o transporte mucociliar nasal por meio do teste de tempo de trânsito da sacarina, a propriedade de superfície do muco nasal por meio do ângulo de contato, a inflamação na cavidade nasal por meio da contagem total e diferencial das células e citocinas em lavado nasal e a inflamação de vias aéreas inferiores por meio do pH do condensado do ar exalado. Observamos neste estudo que os jovens tabagistas >= 2,5 foram mais velhos comparados aos JNT e com os tabagistas < 2,5 (p < 0,01). Comparados com os JNT, os tabagistas >= 2,5 apresentaram maior índice de massa corporal (p=0,036), de frequência cardíaca (p=0,001) e de pressão arterial sistólica (p=0,036). Os tabagistas >= 2,5 apresentaram maior queixa sobre tosse (p=0,05) e secreção nasal escorrendo para a garganta (p=0,016) quando comparados com os JNT e tabagistas < 2,5. Não encontramos diferenças significativas na pontuação total do questionário SNOT-20 (p=0,140), nos valores do teste de função pulmonar e nos valores do ângulo de contato (p=0,803) entre os grupos. O tempo de trânsito da sacarina foi significativamente menor nos jovens tabagistas (5,9 ± 3,1 minutos) quando comparados aos JNT (7,7 ± 4,1 minutos, p=0,033). Na análise do lavado nasal encontramos maior número de células totais em tabagistas < 2,5 (48+-14) e tabagistas >= 2,5 (37+-25) comparados aos JNT (24+-12, p < 0,001). Encontramos também maior número de macrófagos (p=0,001), células ciliadas (p=0,008) e células caliciformes (p=0,004) nos tabagistas < 2,5 e tabagistas >= 2,5 quando comparados aos JNT, e maiores concentrações de mieloperoxidase nos tabagistas < 2,5 comparados aos tabagistas >= 2,5 (p=0,005). Os valores do pH do EBC foram menores em tabagistas >= 2,5 (7,65 ± 0,42) comparados com os tabagistas < 2,5 (7,83 ± 0,26) e com os JNT (7,90 ± 0,21, p=0,038). Por meio de análise de regressão linear, verificamos um efeito dose-dependente significativo do tabagismo sobre a redução dos valores do pH do EBC (r=-0,47, p < 0,001). Concluímos que o tabagismo em jovens tabagistas com idade igual ou inferior a 35 anos induz alterações do transporte mucociliar nasal, inflamação nasal e inflamação em vias aéreas inferiores e que essas alterações estão associadas à carga tabágica / Cigarette smoking is strongly associated with the development of Chronic Obstructive Pulmonary Disease, but few studies that reported functional or inflammatory changes in upper airway in young are found in the literature. We aimed to evaluate the effects of smoking on nasal mucociliary clearance, the mucus surface property and if there is inflammation in the nasal cavity and lower airways in young smokers aged less than 35 years. Of the 200 individuals contacted by telephone, 72 individuals entered in the study: 32 healthy young nonsmokers (YNS) (21 ± 4 years, 29 male) and 40 young smokers, subdivided according to smoking history: less than 2.5 pack-years (< 2.5) (19 ± 2 years, 20 male) and 2.5 pack-years or more ( >= 2.5) (24 ± 5 years, 17 male and three female). We assessed demographic data, clinical data, SNOT-20 questionnaire for symptoms of airway discomfort, the volumes and lung capacities by the pulmonary function test, the nasal mucociliary clearance using the saccharine transit test, the mucus surface property by the contact angle, the inflammation in the nasal cavity by the total and differential count of cells and cytokines in nasal lavage and inflammation of the lower airways by the exhaled breath condensate pH. In this study, we observed that young smokers >= 2.5 were older compared to YNS and smokers < 2.5 (p < 0.01). Compared with YNS, smokers >= 2.5 had higher body mass index (p=0.036), heart rate (p=0.001) and systolic blood pressure (p=.036). Smokers >= 2.5 complained more about cough (p = 0.05) and post-nasal discharge (p=0.016) when compared to YNS and smokers < 2.5. No significant differences were found in the total score of the SNOT-20 (p=0.140), in the pulmonary function test and mucus contact angle (p=0.803) between groups. The saccharine transit time was significantly lower in young smokers (5.9 ± 3.1 minutes) compared to YNS (7.7 ± 4.1 minutes, p=0.033). The number of total cells in nasal lavage fluid were greater in smokers < 2.5 (48±14) and smokers >= 2.5 (37 ± 25) compared to YNS (24 ± 12, p < 0.001). We also found greater number of macrophages (p=0.001), ciliated cells (p=0.008) and goblet cells (p = 0.004) in smokers < 2.5 and smokers >= 2.5 compared to YNS and a higher concentration of myeloperoxidase in smokers < 2.5 compared to smokers >= 2.5 (p=0.005). The EBC pH were lower in smokers >= 2.5 (7.65 ± 0.42) compared with smokers < 2.5 (7.83 ± 0.26) and with YNS (7.90 ± 0.21, p=0.038). Linear regression analysis confirmed a significant dose-dependent effect of smoking in decreasing EBC pH (r= -0.47, p < 0.001). We conclude that cigarette smoking induces changes in nasal mucociliary clearance, nasal inflammation and inflammation in the lower airways in young smokers aged less than 35 years and these changes are associated with smoking history

O tabagismo está fortemente associado ao desenvolvimento da Doença Pulmonar Obstrutiva Crônica, porém poucos estudos que relatam alterações funcionais ou inflamatórias nas vias aéreas superiores em jovens são encontrados na literatura. Nosso objetivo foi investigar as alterações funcionais e inflamatórias nas vias aéreas superiores e inferiores em jovens tabagistas com idade igual ou inferior a 35 anos. Setenta e dois indivíduos participaram do estudo: 32 jovens não tabagistas (JNT) saudáveis (21±4 anos, 29 homens) e 40 jovens tabagistas subdivididos de acordo com a carga tabágica: menor que 2,5 anos-maço (< 2,5) (19±2 anos, 20 homens) e igual ou maior que 2,5 anos-maço (>= 2,5) (24 ± 5 anos, 17 homens e três mulheres). Foram avaliados os dados demográficos, os dados clínicos, os sintomas de desconforto das vias aéreas por meio do questionário SNOT-20, os volumes e capacidades pulmonares por meio do teste de função pulmonar, o transporte mucociliar nasal por meio do teste de tempo de trânsito da sacarina, a propriedade de superfície do muco nasal por meio do ângulo de contato, a inflamação na cavidade nasal por meio da contagem total e diferencial das células e citocinas em lavado nasal e a inflamação de vias aéreas inferiores por meio do pH do condensado do ar exalado. Observamos neste estudo que os jovens tabagistas >= 2,5 foram mais velhos comparados aos JNT e com os tabagistas < 2,5 (p < 0,01). Comparados com os JNT, os tabagistas >= 2,5 apresentaram maior índice de massa corporal (p=0,036), de frequência cardíaca (p=0,001) e de pressão arterial sistólica (p=0,036). Os tabagistas >= 2,5 apresentaram maior queixa sobre tosse (p=0,05) e secreção nasal escorrendo para a garganta (p=0,016) quando comparados com os JNT e tabagistas < 2,5. Não encontramos diferenças significativas na pontuação total do questionário SNOT-20 (p=0,140), nos valores do teste de função pulmonar e nos valores do ângulo de contato (p=0,803) entre os grupos. O tempo de trânsito da sacarina foi significativamente menor nos jovens tabagistas (5,9 ± 3,1 minutos) quando comparados aos JNT (7,7 ± 4,1 minutos, p=0,033). Na análise do lavado nasal encontramos maior número de células totais em tabagistas < 2,5 (48+-14) e tabagistas >= 2,5 (37+-25) comparados aos JNT (24+-12, p < 0,001). Encontramos também maior número de macrófagos (p=0,001), células ciliadas (p=0,008) e células caliciformes (p=0,004) nos tabagistas < 2,5 e tabagistas >= 2,5 quando comparados aos JNT, e maiores concentrações de mieloperoxidase nos tabagistas < 2,5 comparados aos tabagistas >= 2,5 (p=0,005). Os valores do pH do EBC foram menores em tabagistas >= 2,5 (7,65 ± 0,42) comparados com os tabagistas < 2,5 (7,83 ± 0,26) e com os JNT (7,90 ± 0,21, p=0,038). Por meio de análise de regressão linear, verificamos um efeito dose-dependente significativo do tabagismo sobre a redução dos valores do pH do EBC (r=-0,47, p < 0,001). Concluímos que o tabagismo em jovens tabagistas com idade igual ou inferior a 35 anos induz alterações do transporte mucociliar nasal, inflamação nasal e inflamação em vias aéreas inferiores e que essas alterações estão associadas à carga tabágica / Cigarette smoking is strongly associated with the development of Chronic Obstructive Pulmonary Disease, but few studies that reported functional or inflammatory changes in upper airway in young are found in the literature. We aimed to evaluate the effects of smoking on nasal mucociliary clearance, the mucus surface property and if there is inflammation in the nasal cavity and lower airways in young smokers aged less than 35 years. Of the 200 individuals contacted by telephone, 72 individuals entered in the study: 32 healthy young nonsmokers (YNS) (21 ± 4 years, 29 male) and 40 young smokers, subdivided according to smoking history: less than 2.5 pack-years (< 2.5) (19 ± 2 years, 20 male) and 2.5 pack-years or more ( >= 2.5) (24 ± 5 years, 17 male and three female). We assessed demographic data, clinical data, SNOT-20 questionnaire for symptoms of airway discomfort, the volumes and lung capacities by the pulmonary function test, the nasal mucociliary clearance using the saccharine transit test, the mucus surface property by the contact angle, the inflammation in the nasal cavity by the total and differential count of cells and cytokines in nasal lavage and inflammation of the lower airways by the exhaled breath condensate pH. In this study, we observed that young smokers >= 2.5 were older compared to YNS and smokers < 2.5 (p < 0.01). Compared with YNS, smokers >= 2.5 had higher body mass index (p=0.036), heart rate (p=0.001) and systolic blood pressure (p=.036). Smokers >= 2.5 complained more about cough (p = 0.05) and post-nasal discharge (p=0.016) when compared to YNS and smokers < 2.5. No significant differences were found in the total score of the SNOT-20 (p=0.140), in the pulmonary function test and mucus contact angle (p=0.803) between groups. The saccharine transit time was significantly lower in young smokers (5.9 ± 3.1 minutes) compared to YNS (7.7 ± 4.1 minutes, p=0.033). The number of total cells in nasal lavage fluid were greater in smokers < 2.5 (48±14) and smokers >= 2.5 (37 ± 25) compared to YNS (24 ± 12, p < 0.001). We also found greater number of macrophages (p=0.001), ciliated cells (p=0.008) and goblet cells (p = 0.004) in smokers < 2.5 and smokers >= 2.5 compared to YNS and a higher concentration of myeloperoxidase in smokers < 2.5 compared to smokers >= 2.5 (p=0.005). The EBC pH were lower in smokers >= 2.5 (7.65 ± 0.42) compared with smokers < 2.5 (7.83 ± 0.26) and with YNS (7.90 ± 0.21, p=0.038). Linear regression analysis confirmed a significant dose-dependent effect of smoking in decreasing EBC pH (r= -0.47, p < 0.001). We conclude that cigarette smoking induces changes in nasal mucociliary clearance, nasal inflammation and inflammation in the lower airways in young smokers aged less than 35 years and these changes are associated with smoking history

Although public transit bus accounts for only a small percentage of the mode share for transportation in Florida, the annual passenger miles were over 1 billion with over 200 million passenger trips in 2005. These numbers warrant close attention to be paid to the safety of public transit vehicles. Despite the relatively low occurrence of fatalities and bus crashes, each crash of a high occupancy vehicle such as a public transit bus could expose more people to injury than a private automobile crash. Bus crashes also have a significant impact on the automobiles that are involved. Since a high percentage of bus crashes in Florida are caused by rear-end collisions with private automobiles, improving the signage and lighting that will allow buses to move back into traffic safely is very important for bus safety and operations. This paper uses bus operator surveys, crash data, and field studies to develop recommendations for lighting and signage on the back of the bus, roadway signs and revised Florida legislations. Improved signage and lighting will help the bus move back into traffic safely, decrease bus delay and improve bus operations however it must be accompanied by laws and law enforcement.

<p>Bikunin, a chondroitin sulfate-containing protein of 25 kDa, has protease inhibitory activity and occurs in the plasma in free and complexed form. In inter-α-inhibitor (IαI) and pre-a-inhibitor (PαI) it is covalently linked through its chondroitin sulfate (CS) chain to two or one other polypeptide of about 80 kDa – heavy chains 1 and 2 (H1, H2) and heavy chain 3 (H3) – respectively. Bikunin and the heavy chains are synthesized as precursors, which are proteolytically cleaved and assembled into IαI and PαI in the secretory pathway. The C-terminal extension (CTX) of the heavy chains seems to mediate its own cleavage and theassembly of the complexes. The heavy chains of the IαI family become transferred to hyaluronan during ovulation and inflammation.</p><p>In this thesis, the biosynthesis of PαI, the plasma clearance of bikunin and the binding of IαI to collagen were studied. We found that in H3, a short segment on the N-terminal side of the CTX cleavage site is required for cleavage. Furthermore, the H3 could become linked to free CS chains primed by a xyloside, showing that the bikunin protein core is not needed for coupling. We also identified His649 as a residue essential for coupling, but not for cleavage. </p><p>Bikunin labelled with a residualizing agent, 125I-tyramine cellobiose, was injected into mice to identify tissues involved in its uptake. Half of the radioactivity was recovered in the kidneys, 10% in the liver, and the rest distributed in other tissues. We determined the half-life of bikunin in rat plasma using two independent methods: injection of 125I-bikunin, or hepatectomy followed by assessing the rate of disappearance of endogenous bikunin. Both methods yielded half-time values of 5-7 minutes. Removal of the CS chain did not affect the clearance rate of bikunin.</p><p>IαI and its heavy chains were found to bind to collagen with dissociation constants greater than 2 μM and 0.4-0.6 μM, respectively and this binding was independent of divalent metal ions. We suggest that the interaction of IαI with collagen may play a modulatory role in cell migration or in remodelling of the extracellular matrix.</p>

Bikunin, a chondroitin sulfate-containing protein of 25 kDa, has protease inhibitory activity and occurs in the plasma in free and complexed form. In inter-α-inhibitor (IαI) and pre-a-inhibitor (PαI) it is covalently linked through its chondroitin sulfate (CS) chain to two or one other polypeptide of about 80 kDa – heavy chains 1 and 2 (H1, H2) and heavy chain 3 (H3) – respectively. Bikunin and the heavy chains are synthesized as precursors, which are proteolytically cleaved and assembled into IαI and PαI in the secretory pathway. The C-terminal extension (CTX) of the heavy chains seems to mediate its own cleavage and theassembly of the complexes. The heavy chains of the IαI family become transferred to hyaluronan during ovulation and inflammation. In this thesis, the biosynthesis of PαI, the plasma clearance of bikunin and the binding of IαI to collagen were studied. We found that in H3, a short segment on the N-terminal side of the CTX cleavage site is required for cleavage. Furthermore, the H3 could become linked to free CS chains primed by a xyloside, showing that the bikunin protein core is not needed for coupling. We also identified His649 as a residue essential for coupling, but not for cleavage. Bikunin labelled with a residualizing agent, 125I-tyramine cellobiose, was injected into mice to identify tissues involved in its uptake. Half of the radioactivity was recovered in the kidneys, 10% in the liver, and the rest distributed in other tissues. We determined the half-life of bikunin in rat plasma using two independent methods: injection of 125I-bikunin, or hepatectomy followed by assessing the rate of disappearance of endogenous bikunin. Both methods yielded half-time values of 5-7 minutes. Removal of the CS chain did not affect the clearance rate of bikunin. IαI and its heavy chains were found to bind to collagen with dissociation constants greater than 2 μM and 0.4-0.6 μM, respectively and this binding was independent of divalent metal ions. We suggest that the interaction of IαI with collagen may play a modulatory role in cell migration or in remodelling of the extracellular matrix.

In the present study, possible scale effects on the formation of air-entraining vortices at horizontal intakes are studied experimentally. Basic dimensionless parameters that govern the onset of vortices at a horizontal intake in a model and a prototype are stated by dimensional analysis. Series of experiments are conducted on four intake pipes of different diameters located in a large reservoir. The relationship of critical submergence ratio with other dimensionless parameters is considered for a given Froude number and it is found out that the critical submergence ratio is affected by model length scale ratio and its natural result of side-wall clearance ratio and Reynolds number differences between model and prototypes. It is observed that, side-wall clearance ratio is not effective on the critical submergence ratio after it exceeds about 6. In addition to this, Reynolds number limit, beyond which viscous forces do not affect the vortex flow, is found out to be increasing with the increase in Froude number. An empirical relationship, which gives the critical submergence ratio as a function of Froude number, side-wall clearance ratio and Reynolds number is obtained by using data collected in the experiments.